Treatment of myelosuppression

ABSTRACT

Methods are presented for attenuating myelosuppressive side effects of treatment regimens, promoting thrombopoiesis and neutrophil production, and increasing efficacy of treatment regimens, by administering PF4-interacting heparinoids.

1. CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.15/001,773, filed Jan. 20, 2016, which is a continuation of U.S.application Ser. No. 13/801,990, now U.S. Pat. No. 9,271,999, filed Mar.13, 2013, which claims the benefit of U.S. Provisional Application Nos.61/724,836, filed Nov. 9, 2012; 61/702,207, filed Sep. 17, 2012;61/678,053, filed Jul. 31, 2012; 61/668,709, filed Jul. 6, 2012;61/664,611, filed Jun. 26, 2012; 61/653,362, filed May 30, 2012;61/648,043, filed May 16, 2012; 61/644,623, filed May 9, 2012; and61/644,556, filed May 9, 2012, the contents of all of which areincorporated herein in their entireties by reference thereto.

2. BACKGROUND

Platelets play a critical role in the blood clotting mechanism.Depletion of platelets below a certain level results inthrombocytopenia, which can be triggered by a number of clinicalconditions and disorders and can range from mild to life-threatening.

Thombocytopenia can be triggered by diseases and conditions affectingthe bone marrow, where platelet precursors arise before entering thebloodstream; by diseases and conditions affecting the liver, whichproduces thrombopoietin, the hormone that stimulates the production ofplatelets; by sequestration of platelets; by increased destruction ofplatelets; and by a variety of other causes. In particular,thrombocytopenia is a common side effect of certain treatment regimens,such as cancer treatment regimens involving antineoplastic agents.Chemotherapy-induced or radiation-induced thrombocytopenia can result indelays in treatment and/or compel reductions in treatment dose, which inturn can result in reduced efficacy of the treatment.

Because severe thrombocytopenia puts a patient at risk of uncontrolledhemorrhage, development of safe and effective treatments forthrombocytopenia is highly desirable. In spite of the clear need forsuch treatments, however, very few such treatments exist. Attempts todevelop a recombinant form of human thrombopoietin have provedunsuccessful. While a recombinant human thrombopoietin showed earlypromise, it showed a tendency to induce the development ofauto-antibodies when tested in patients. Currently, standard therapy ofthrombocytopenia, such as immune-mediated thrombocytopenia, can includetreatment with corticosteroids, rituximab, and/or thrombopoietinreceptor agonists, splenectomy, and platelet transfusions. However, eachhas drawbacks, including incomplete response, development of sideeffects to treatment, and risks attendant to any form of surgery. Forchemotherapy-induced thrombocytopenia, only one therapeutic agent,interleukin-11, has proven sufficiently effective to merit regulatoryapproval, but is rarely prescribed by physicians because of the severityof its side effects. For radiation-induced thrombocytopenia, there is noapproved therapeutic agent that attenuates the thrombocytopenia. Thus,there remains a significant need for agents that attenuatethrombocytopenias of varying etiology, including immune-mediatedthrombocytopenias, drug-induced thrombocytopenias, especiallychemotherapy-induced thrombocytopenia, and radiation-inducedthrombocytopenia.

Neutrophils, also called polymorphonuclear leukocytes, are the mostnumerous of the blood cells known as granulocytes. Neutrophils, likeother blood cells, are produced by the bone marrow. Neutrophils are animportant component of natural immunity. When neutrophil levels fallbelow normal, a condition called neutropenia occurs, increasing the riskof infection. Neutropenia can arise from a number of different causes,ranging from congenital defects to viral infections, but a context inwhich neutropenia frequently occurs is as a side effect of a treatmentregimen. Neutropenia is a common side effect in patients being treatedfor cancer with antineoplastic agents, putting patients at risk ofdeveloping serious and even life-threatening infections, and forcingdelays in treatment and/or compelling reduction in treatment dose,resulting in reduced efficacy.

A variety of agents and therapies have been tested to combatneutropenia, with varying degrees of success. Administration ofglucocorticoids, androgenic steroids, and vitamins to stimulate bonemarrow to produce more neutrophils has not proved successful. Atpresent, only two agents—granulocyte colony-stimulating factor (G-CSF)and granulocyte-macrophage colony-stimulating factor (GM-CSF)—are widelyused to treat patients with severe neutropenia, most often afterintensive cancer chemotherapy and/or bone marrow transplantation. Theseagents exhibit adverse effects such as bone pain, abnormalities of liverdysfunctions and pleural and pericardial effusions. Thus, there is aneed for compounds that are safe and effective for treating neutropeniaand promoting neutrophil production.

3. SUMMARY

In a first aspect, methods are provided for attenuating amyelosuppressive side effect of a patient treatment regimen. The methodscomprise adjunctively administering to the patient a therapeuticallyeffective amount of a platelet factor 4-interacting heparinoid(hereinafter “PF4-interacting heparinoid”). As provided herein, amyelosuppressive side effect is the occurrence of thrombocytopeniaand/or neutropenia, and a patient treatment regimen having amyelosuppressive side effect is a treatment regimen that induces, as aside effect, one or both of thrombocytopenia and neutropenia. In variousembodiments, this aspect provides uses of a PF4-interacting heparinoid,e.g., ODSH, in the attenuation of a myelosuppressive side effect of apatient treatment regimen.

Numerous patient treatment regimens have myelosuppressive side effects,including antineoplastic treatment regimens, such as chemotherapy andradiation therapy, antibody therapy, including treatment regimens usedto treat cancer and/or auto-immune diseases, and transplant procedures,such as bone marrow or stem cell transplant. In certain embodiments, thepatient treatment regimen comprises chemotherapy and/or radiationtherapy and/or antibody therapy. In an exemplary embodiment, the patienttreatment regimen is a chemotherapeutic regimen comprising gemcitabineand nab-paclitaxel. In an exemplary embodiment, the patient treatmentregimen is a chemotherapeutic regimen comprising ifosfamide,carboplatin, and etoposide, optionally including rituximab. In certainembodiments, the patient treatment regimen comprises one or moreregimens suitable for the treatment of subjects diagnosed with acutemyelogenous or myeloid leukemia (“AML”). In an exemplary embodiment, aregimen suitable for the treatment of subjects diagnosed with AMLcomprises a chemotherapy regimen suitable for inducing remission of AML(known in the art as induction chemotherapy), a chemotherapy regimen forpreventing remission of AML (known in the art as consolidationchemotherapy), or both induction and consolidation chemotherapy.Optionally, a regimen suitable for the treatment of subjects diagnosedwith AML can also comprise one or more non-chemotherapy-based regimensfor preventing remission of AML, which can be used instead of or incombination with consolidation chemotherapy. Thesenon-chemotherapy-based regimens include stem cell transplant, such asallogeneic stem cell transplant, and immunotherapy. Further patienttreatment regimens are described in Section 5.1.1.

In various embodiments, the methods are useful for treating subjectsdiagnosed with cancer, for example pancreatic cancer, solid tumorsincluding osteosarcoma, neuroblastoma, or AML. The subject being treatedcan be an adult or a pediatric patient. In some embodiments, the subjectis diagnosed with a cancer in which PF4 levels are elevated either inplatelets or in blood (referred to hereinafter as “PF4-positivecancer”). Examples of PF4-positive cancers include pancreatic cancer andcolorectal cancer. Further suitable subjects are described in Section5.1.2.

In some embodiments, the methods may further comprise adjunctiveadministration of one or more additional agent or therapy that ispro-thrombopoietic, anti-thrombocytopenic, anti-neutropenic,pro-granulopoietic, and/or anticoagulant. Suitable agents and therapiesfor further adjunctive administration are described herein at Section5.1.3. In some embodiments, two or more agents and/or therapies areadministered. The two or more agents can have the same activity (e.g.,anti-thrombocytopenic), different activity (e.g., a first agent ispro-thrombopoietic and a second agent is anti-neutropenic), oroverlapping activity (e.g., a first agent is pro-granulopoietic andanticoagulant and a second agent is anti-coagulant).

The PF4-interacting heparinoid, and any adjunctively administeredadditional agent(s) or therapy, can be administered concurrently,sequentially, or separately from administration of the patient treatmentregimen. Suitable routes and modes of administration are provided belowin Section 5.8.

In a second aspect, methods are provided for promoting thrombopoiesis ina subject. The methods comprise administering an effective amount of aPF4-interacting heparinoid to the subject. The subject can bethrombocytopenic or non-thrombocytopenic, as described below in Section5.2. In various embodiments, the methods comprise treatingmyelosuppression caused by a disease or condition that reduces plateletcount in a subject. In an exemplary embodiment, the subject is diagnosedwith systemic inflammatory response syndrome (SIRS), sepsis, orsepticemia. Optionally, the subject has elevated serum or plasma levelof PF4. In some embodiments, the methods further comprise adjunctiveadministration of one or more additional agent or therapy that ispro-thrombopoietic, anti-thrombocytopenic, anti-neutropenic,pro-granulopoietic, and/or anticoagulant. Suitable agents and therapiesfor further adjunctive administration are described herein at Section5.2.1.

In a third aspect, methods are provided for promoting neutrophilproduction in a subject. The methods comprise administering an effectiveamount of a PF4-interacting heparinoid to the subject. The subject canbe neutropenic or non-neutropenic, as described below in Section 5.3. Invarious embodiments, the methods comprise treating myelosuppressioncaused by a disease or condition that reduces neutrophil count in asubject. Optionally, the subject has elevated serum or plasma level ofPF4. In some embodiments, the methods further comprise adjunctiveadministration of one or more additional agent or therapy that ispro-thrombopoietic, anti-thrombocytopenic, anti-neutropenic,pro-granulopoietic, and/or anticoagulant. Suitable agents and therapiesfor further adjunctive administration are described herein at Section5.3.1.

In a fourth aspect, methods are provided for increasing the efficacy ofa patient treatment regimen having a myelosuppressive side effect. Themethods comprise administering a therapeutically effective amount of aPF4-interacting heparinoid to the subject patient as an adjunct to thepatient treatment regimen having a myelosuppressive side effect, withoutreducing the dose and/or dosage frequency of the patient treatmentregimen following a reference treatment administration or treatmentcycle.

In some embodiments, the method further comprises administering a dosehigher than is typically used for such administration or cycle in theabsence of adjunctive administration of a PF4-interacting heparinoid.

In certain embodiments, the methods further comprise determining aninitial platelet count in a blood sample from a patient andadministering an amount of a PF4-interacting heparinoid effective toraise the patient's platelet count above a threshold level below whichtherapy with patient treatment regimen having a myelosuppressive sideeffect is contraindicated. In various embodiments, an amount of aPF4-interacting heparinoid is administered sufficient to maintainplatelet levels above levels that indicate grade 3 (severe) or grade 4(life-threatening) thrombocytopenia. Optionally, the methods can furthercomprise administering adjunctively to the PF4-interacting heparinoidone or more agents or therapies that is anti-thrombocytopenic,anti-neutropenic, anticoagulant, or has some other therapeutic activity.In some embodiments, the methods comprise a further step ofadministering a patient treatment regimen having a myelosuppressive sideeffect to the patient whose platelet count is above a threshold levelthat contraindicates such therapy. Optionally, the dose amount and/orfrequency of the patient treatment regimen can be increased.

In certain embodiments, the methods comprise determining an initialneutrophil count in a blood sample from a patient and administering anamount of a PF4-interacting heparinoid effective to raise the patient'sneutrophil count above a threshold level below which therapy withpatient treatment regimen having a myelosuppressive side effect iscontraindicated. In various embodiments, an amount of a PF4-interactingheparinoid is administered sufficient to maintain neutrophil levelsabove levels that indicate grade 3 or grade 4 neutropenia, i.e., aboveabout 1000 neutrophils/of blood and above about 500 neutrophils/μl ofblood, respectively. Optionally, the methods can further compriseadministering adjunctively to the PF4-interacting heparinoid one or moreagents or therapies that is anti-neutropenic, anti-thrombocytopenic,anticoagulant, or has some other therapeutic activity. In someembodiments, the methods comprise a further step of administering apatient treatment regimen having a myelosuppressive side effect to thepatient whose neutrophil count is above a level that contraindicatessuch therapy. Optionally, the dose amount and/or frequency of thepatient treatment regimen can be increased.

The PF4-interacting heparinoids of the present disclosure areheparinoids that are capable of interacting with PF4 and counteractingPF4′s ability to suppress production of platelets and neutrophils.PF4-interacting heparinoids bind to PF4 and/or compete with PF4 forbinding to progenitor cells in the myeloid cell lineage, e.g.,megakaryocytes. Preferably, PF4-interacting heparinoids have an averagemolecular weight above about 8 kDa, such as an average molecular weightbetween about 8 kDa and about 15 kDa, more preferably between about 11kDa and 13 kDa. The PF4-interacting heparinoid is preferably partiallydesulfated. In some embodiments, the PF4-interacting heparinoid issubstantially sulfated at the 6-O and/or the N position. An exemplaryPF4-interacting heparinoid, which is suitable for use in the methodsdescribed herein, is substantially 2-O, 3-O desulfated, referred toherein as ODSH. See also Section 5.6.

The present disclosure further provides pharmaceutical compositions andunit dosage forms comprising PF4-interacting heparinoids, suitable foruse in the methods described herein, either alone or adjunctive to apatient treatment regimen and/or one or more additional agent ortherapy. The pharmaceutical compositions may be prepared for parenteraladministration, such as intravenous or subcutaneous administration. Forintravenous administration, pharmaceutical compositions can beformulated for administration as a bolus or as a continuous infusion.

Pharmaceutical compositions for use in the methods disclosed hereincomprise an amount of a PF4-interacting heparinoid, as described belowin Section 5.9, sufficient to allow effective doses to be administered.

In some embodiments, pharmaceutical compositions of PF4-interactingheparinoid are suitable for intravenous administration at doses rangingfrom about 0.1 mg/kg/hr to about 2.5 mg/kg/hr for infusions and fromabout 1 mg/kg to about 25 mg/kg for bolus doses. In some embodiments,pharmaceutical compositions of PF4-interacting heparinoid are suitablefor subcutaneous administration and are formulated for administration atdoses ranging from about 25 mg to about 400 mg, in volumes of 2.0 ml orless per injection site.

4. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 provides a graph illustrating the effect of 8 different treatmentregimens on tumor weight in an in vivo murine xenograft model of humanpancreatic cancer, as described further in Example 1 and at Table 1:vehicle control (Group 1, ●); ODSH alone (Group 2, ∘);oxaliplatin/gemcitabine/nab-paclitaxel (Group 3, ▪); gemcitabine alone(Group 4, □); oxaliplatin/gemcitabine/nab-paclitaxel with ODSH (Group 5,▴); gemcitabine with ODSH (Group 6, Δ); oxaliplatin/gemcitabine (Group7, ×); and oxaliplatin/gemcitabine with ODSH (Group 8, ⋆).

FIG. 2 provides a graph illustrating the effect on tumor weight of asubset of the treatment regimens used in Example 1 and shown in FIG. 1:vehicle control (Group 1, ●); ODSH alone (Group 2, ∘); gemcitabine alone(Group 4, □); and gemcitabine with ODSH (Group 6, Δ).

FIG. 3 provides a graph illustrating the effect on body weight of the 8different regimens used in Example 1: vehicle control (Group 1, ●); ODSHalone (Group 2, ∘); oxaliplatin/gemcitabine/nab-paclitaxel (Group 3, ▪);gemcitabine alone (Group 4, □); oxaliplatin/gemcitabine/nab-paclitaxelwith ODSH (Group 5, ▴); gemcitabine with ODSH (Group 6, Δ);oxaliplatin/gemcitabine (Group 7, ×); and oxaliplatin/gemcitabine withODSH (Group 8, ⋆).

FIG. 4 provides a graph illustrating the effect on tumor weight of 4different treatment regimens in an in vivo murine xenograft model ofhuman ovarian cancer, as described further in Example 2: vehicle control(Group 1, ●); ODSH alone (Group 2, ∘); carboplatin (Group 3, ▪); andcarboplatin with ODSH (Group 4, □).

FIG. 5 provides a graph illustrating the effect on tumor weight of thetreatment regimens shown in FIG. 4, for days 1-21 of the study.

FIG. 6 provides a graph illustrating the effect on mouse body weight ofthe 4 different regimens used in Example 2: vehicle control (Group 1,●); ODSH alone (Group 2, ∘); carboplatin (Group 3, ▪); and carboplatinwith ODSH (Group 4, □), as described further in Example 2.

FIG. 7 provides a graph illustrating the effect on mouse body weight ofthe treatment regimens shown in FIG. 6 for days 1-21.

FIG. 8 provides a chart of the platelet count (in ×10³ platelets/μL) ofpatients with metastatic pancreatic cancer entered in the clinical trialdescribed in Example 3, as measured in samples taken on day 1, day 8,and day 15 of each chemotherapy cycle as indicated (C1D1=cycle 1, day 1;C2D8=cycle 2, day 8, etc.). Horizontal lines mark the lower limit ofnormal platelet count (LLN) and the lower limit (LL) of the indicatedgrades of thrombocytopenia.

FIG. 9 provides a chart of the neutrophil count (in ×10³ neutrophils/μL)measured in samples taken from the same individuals described in FIG. 8,on day 1, day 8, and day 15 of each chemotherapy cycle as indicated(C1D1=cycle 1, day 1; C2D8=cycle 2, day 8, etc.). Horizontal lines markthe lower limit of normal neutrophil count (LLN) and the lower limit(LL) of the indicated grades of neutropenia.

FIG. 10A-10B provide mean and median platelet counts (FIG. 10A) and meanand median absolute neutrophil counts (FIG. 10B) for all samples at theindicated days in each of the indicated cycles.

FIGS. 11A-F provides charts of the size of pancreatic and metastaticlesions in specific patients enrolled in the clinical trial described inExample 3 who have stable disease and who are receiving adjunctiveadministration of ODSH and chemotherapeutic agents. FIGS. 11A shows thesize of two pulmonary metastases in patient 2001 at baseline and at theend of cycle 4. FIGS. 11B-C show tumor size of metastatic lesions in theliver and lymph nodes at the end of treatment cycle 2 (FIG. 11B) andlesions in the pancreas, liver and lymph nodes at the end of cycle 5(FIG. 11C) relative to the start of treatment (baseline) for patient6002. FIG. 11D shows the size of two pulmonary metastases in patient6003 at baseline and at the end of cycle 6. FIG. 11E shows the size ofpancreatic tumors and a metastatic liver tumor in patient 6006 atbaseline and at the end of cycle 4. FIG. 11F shows the size of apancreatic tumor in patient 8001 at baseline and at the end of cycle 2.

FIGS. 12A-F provide charts showing the tumor response of patientsreceiving adjunctive administration of ODSH and chemotherapy. FIG. 12Aprovides a chart summarizing sites of metastatic disease before thestart of chemotherapy, levels of CA19-9 at baseline and after severalchemotherapy cycles, and tumor response for each indicated patient. FIG.12B-12F provides charts of the size of tumors for patients 6004, 6007,7001, 7002, and 9001, showing a partial response at the end of treatmentcycle 4 or 5, relative to baseline.

FIGS. 13A-B provide charts showing platelet counts at days 1 and 15 ofindicated cycles for patients treated with gemcitabine, nab-paclitaxel,and ODSH (“ODSH arm patients”) and patients treated with gemcitabine andnab-paclitaxel (“Control arm patients”). FIG. 13A provides a chartshowing platelet counts, at day 1 of cycle 1 (before any chemotherapy)and at day 15 of cycle 1 (after two doses of chemotherapy) in 5 ODSH armpatients and 5 Control arm patients. Median and mean platelet counts forthe control arm and the ODSH arm at days 1 and 15 are also shown. FIG.13B provides a chart showing median platelet counts at days 1 and 15 ofcycles 1 through 6 for Control arm patients and ODSH arm patients.

5. DETAILED DESCRIPTION

It has been discovered that heparinoids that are capable of interactingwith platelet factor 4 (hereinafter, “PF4-interacting heparinoids”) canattenuate thrombocytopenia and neutropenia of various etiologies. It hasfurther been found that PF4-interacting heparinoids induce or disinhibitthrombopoiesis and granulopoiesis. Without intending to be bound bytheory, it is thought that these effects are mediated by the ability ofsuch heparinoids to reduce PF4 levels and/or counteract a suppressiveeffect of PF4 on megakaryopoiesis and granulopoiesis.

5.1. Methods of Attenuating Myelosuppressive Side Effects of TreatmentRegimens

As described in Example 3 below, patients diagnosed with metastaticpancreatic cancer who were treated adjunctively with an exemplaryPF4-interacting heparinoid, referred to herein as ODSH (a heparinoidthat is substantially desulfated at 2-O and 3-O positions, furtherdescribed in Section 5.6), had increased platelet counts at the end of afirst 4 week cycle of a chemotherapy regimen that is known to have asubstantial myelosuppressive side effect. These effects continued insuccessive cycles of treatment and the results demonstrated thatadjunctively administered ODSH attenuates thrombocytopenia andneutropenia in patients receiving a chemotherapy treatment regimenhaving myelosuppressive side effects.

Thus, in a first aspect, methods are provided for attenuating amyelosuppressive side effect of a patient treatment regimen. The methodscomprise administering a therapeutically effective amount of aPF4-interacting heparinoid to the subject patient as an adjunct to thepatient treatment regimen having myelosuppressive side effect. Thus,provided herein are uses of a PF4-interacting heparinoid, optionallyODSH, in the attenuation of a myelosuppressive side effect of a patienttreatment regimen, as discussed further herein. The phrases “adjunctiveadministration”, “adjunctively administering” or “administeringadjunctive to” are used interchangeably herein to mean administering aPF4-interacting heparinoid in therapeutically effective temporalproximity to the treatment regimen that has a myelosuppressive sideeffect. By adjunctively administering a PF4-interacting heparinoid topatients receiving treatment regimens having a myelosuppressive sideeffect, either alone or in combination with other adjunctive agent(s) ortherapy, Applicant has discovered that it is possible to attenuate themyelosuppressive side effect(s) of such treatment regimens.

PF4-interacting heparinoids suitable for use in the methods aredescribed below in Section 5.6. In an exemplary embodiment, thePF4-interacting heparinoid is ODSH. Suitable modes of administration anddosing regimens are described further below, in Section 5.8. Effectivedosages, and therapeutically effective amounts, of PF4-interactingheparinoid are described further below, in Section 5.9.

5.1.1. Treatment Regimens with Myelosuppressive Side Effects

As used herein, a myelosuppressive side effect is the occurrence ofthrombocytopenia and/or neutropenia. Thus, in various embodiments, thetreatment regimen, as a side effect, causes patients to developthrombocytopenia (low platelet count), neutropenia (low neutrophilcount), or a combination of thrombocytopenia and neutropenia. Suchpatient treatment regimens are also referred to herein asmyelosuppressive treatment regimens.

In certain embodiments, the treatment regimen causes thrombocytopenia.In various embodiments, the treatment regimen causes platelet counts inblood to be less than about 150,000 platelets per μl of blood. Inparticular embodiments, the treatment regimen causes the patient to haveplatelet counts ranging from about 150,000 to about 75,000 platelets perμl of blood, corresponding to mild or grade 1 thrombocytopenia; plateletcounts ranging from less than about 75,000 to about 50,000 platelets perμl of blood, corresponding to moderate or grade 2 thrombocytopenia;platelet counts ranging from less than about 50,000 to about 25,000platelets per μl of blood, corresponding to severe or grade 3thrombocytopenia; and platelet counts of less than about 25,000platelets per μl of blood, corresponding to life-threatening or grade 4thrombocytopenia. Thus, in a variety of embodiments, the patienttreatment regimen induces, as a side effect, mild, moderate, severe, orlife-threatening thrombocytopenia.

In certain embodiments, the treatment regimen causes neutropenia. Invarious embodiments, the treatment regimen causes patients to haveabsolute neutrophil counts in blood of less than about 2000 neutrophilsper μl of blood. In particular embodiments, the treatment regimen causesthe patient to have neutrophil counts ranging from about 2000 to about1500 neutrophils per μl of blood, corresponding to mild or grade 1neutropenia; absolute neutrophil counts ranging from less than about1500 to about 1000 neutrophils per μl of blood, corresponding tomoderate or grade 2 neutropenia; absolute neutrophil counts ranging fromless than about 1000 to about 500 neutrophils per μl of blood,corresponding to severe or grade 3 neutropenia; and absolute neutrophilcounts of less than about 500 neutrophils per μl of blood, correspondingto life-threatening or grade 4 neutropenia. Thus, in a variety ofembodiments, the patient treatment regimen induces, as a side effect,mild, moderate, severe, or life-threatening neutropenia.

In a variety of embodiments, the patient treatment regimen is anantineoplastic treatment regimen. In certain embodiments, theantineoplastic treatment regimen is chemotherapy. In certainembodiments, the antineoplastic treatment regimen is radiation therapy.

In chemotherapy embodiments, the patient treatment regimen includesadministration of one or more chemotherapeutic agent(s).

In exemplary embodiments, at least one of the one or morechemotherapeutic agents is selected from the group consisting of: folateantagonists, including methotrexate and pemetrexed; purine antagonists,including cladribine, clofarabine, fludarabine, 6-mercaptopurine,nelarabine, pentostatin; pyrimidine antagonists, including capecitabine,cytarabine, 5-fluorouracil, gemcitabine, hydroxyurea; biologic responsemodifiers, including interferon-alfa; bleomycin; DNA alkylating agents,including nitrosureas, carmustine, lomustine; DNA cross-linking drugsand alkylating agents, including bendamustine, chlorambucil,cyclophosphamide, ifosfamide, mechlorethamine (nitrogen mustard),melphalan, dacarbazine, temozolomide, procarbazine; asparaginase;antibiotics, including mitomycin; platinum complexes, includingcarboplatin, cisplatin, oxaliplatin; proteosome inhibitors, includingbortezomib; spindle poisons, such as the taxanes (including docetaxel,paclitaxel, nab-paclitaxel (Abraxane®)) and the vincas (includingvinblastine, vincristine, vinorelbine); topoisomerase inhibitors, suchas the anthracyclines (including daunorubicin, daunomycin, doxorubicin,epirubicin), the camptothecines, (including irinotecan, topotecan), thepodophyllotoxins (including etoposide, teniposide and mitoxantrone);tyrosine kinase inhibitors, (including erlotinib (Tarceva), gefitinib,imatinib, lapatinib, pazopanib, sorafenib, sunitinib); and ifosfamide.

In various embodiments, one or more other chemotherapeutic agents areused.

In certain exemplary embodiments, the myelosuppresive chemotherapeutictreatment regimen includes administration of a taxane, such asdocetaxel, or a taxol, such as paclitaxel (e.g., nab-paclitaxel,Abraxane®) in combination with one or more additional chemotherapeuticagent(s), including but not limited to any of the agents describedabove. In some embodiments, the patient treatment regimen includesadministration of a taxane, such as docetaxel, or a taxol, such aspaclitaxel (e.g., nab-paclitaxel, Abraxane®) in combination with one ormore of a folate, purine, or pyrimidine antagonist, a DNA alkylatingagent, a platinum complex, a vinca, an anthracycline, a camptothecine, apodophyllotoxin, and/or a tyrosine kinse inhibitor. In specificembodiments, the patient treatment regimen includes administration of ataxane, such as docetaxel, paclitaxel (e.g., nab-paclitaxel, Abraxane®)in combination with one or more agent selected from: gemcitabine,vinorelbine, carboplatin, cisplatin, oxaliplatin, temozolomide, andmifepristone. In exemplary embodiments, two or more chemotherapeuticagents are administered, the two or more chemotherapeutic agentsselected from: cisplatin and etoposide; carboplatin and etoposide;cisplatin and irinotecan; carboplatin and irinotecan; cyclophosphamide,doxorubicin (Adriamycin), and vincristine;cyclophosphamide/doxorubicin/vincristine (known as the CAV regimen);gemcitabine with vinorelbine or paclitaxel or nab-paclitaxel(Abraxane®); gemcitabine or capecitabine with oxaliplatin; cisplatin orcarboplatin plus another chemotherapeutic agent; 5-fluorouracil with oneor more of leuvocorin, oxaliplatin, irinotecan.

The myelosuppressive patient treatment regimen, in various embodiments,comprises administration of chemotherapeutic agents according tospecific named regimens. In exemplary embodiments, the patientchemotherapy treatment regimen includes one or more of the followingspecific regimens: 5FU Mayo, 5FU Roswell Park, LVFU2, FOLFOX4, FOLFOX6,bFOL, FUFOX, IFL, XELOX, XELIRI, and CAPIRI, which are described infurther detail in Chau et al., 2009, Br. J. Cancer 100:1704-19; andField et al., 2007, World J. Gastroenterol. 13:3806-15, both of whichare incorporated herein by reference. Another specific named regimen isCHOP, combining cyclophosphamide, hydroxydaunorubicin (or doxorubicin oradriamycin), vincristine (or oncovin), and prednisone or prednisolone,generally used to treat patients with non-Hodgkin's lymphoma. In someembodiments, e.g., where the patient being treated has a history ofcardiovascular disease, doxorubicin is omitted from the regimen, whichis then referred to as COP or CVP. Optionally, the CHOP regimen can befurther combined with rituximab (Rituxan) and is then referred to asR-CHOP or CHOP-R. Other combinations are also possible. Another specificnamed regimen is ICE, combining chemotherapeutic agents ifosfamide,carboplatin, and etoposide. See Habermann, 2012, Hematology 17 Suppl1:S93-7, which is incorporated herein by reference.

Antineoplastic patient treatment regimens that include radiation therapyhave also been shown to have myelosuppressive side effects, sometimesreferred to as radiation-induced thrombocytopenia and radiation-inducedneutropenia. In various radiation embodiments, the patient treatmentregimen includes radiation therapy selected from radiation therapy withx-rays, gamma rays, neutrons, protons, and other sources, external beamradiation therapy, and internal radiation therapy, such asbrachytherapy.

Patient treatment regimens in which one or more antibodies having acytotoxic effect are administered, referred to herein as antibodytherapy, may also have myelosuppressive side effects that are usefullytreated by adjunctive administration of a PF4-interacting heparinoidaccording to the methods described herein. Thus, in certain embodiments,the treatment regimen having myelosuppressive side effects comprisesantibody therapy. In some embodiments, the antibody therapy includes oneor more antibodies conjugated to a toxin, where the antibody binds toand/or is internalized by a target tumor cell and the toxin kills thecell. In exemplary embodiments, the patient treatment regimen includesadministration of one or more antibodies having a myelosuppressive sideeffect, such as abciximab (ReoPro), rituximab (Rituxan), trastuzumab(Herceptin) conjugated to mertansine (T-DM1), and infliximab (Remicade).In some embodiments, the patient treatment regimen includesadministration of one or more of the following: trastuzumab (Herceptin),cetuximab, bevacizumab (Avastin), tigatuzumab.

In various embodiments, patient treatment regimens that havemyelosuppressive side effects, and that are usefully treated byadjunctive administration of a PF4-interacting heparinoid according tothe methods described herein, include combinations of chemotherapy,radiation therapy and/or antibody therapy. In some embodiments, thepatient treatment regimen comprises chemotherapy, e.g. with one or moreof the agents described herein, and radiation therapy; or chemotherapy,e.g. with one or more of the agents described herein, and antibodytherapy, e.g. with one or more of the antibodies described herein;radiation therapy and antibody therapy, e.g., with one or more of theantibodies described herein; or any two, three, four, five or moreagents or therapies described herein. In exemplary embodiments, such aswhere the patient has non-Hodgkin's lymphoma, the patient treatmentregimen comprises antibody therapy with rituxan and chemotherapy withCHOP (also referred to as R-CHOP), COP, CVP, or ICE (also referred to asR-ICE) regimen. See Habermann, 2012, Hematology 17 Suppl 1:S93-97.

Patient treatment regimens involving transplantation, such as bonemarrow transplant or stem cell transplant, may also havemyelosuppressive side effects. Thus, in some embodiments, the patienttreatment regimen comprises an autologous or allogeneic bone marrow orstem cell transplant.

In a variety of embodiments, the patient treatment regimens includeregimens in which one or more agents with thrombocytopenic side effectsare administered. In exemplary embodiments, the one or more agent with athrombocytopenic side effect is selected from: valproic acid, protonpump inhibitors, interferon (e.g. interferon-alpha), isotretinoin,panobinostat, thiazide diurectics, montelukast sodium (Singulair),quinidine, quinine, gold, sulfonamides, cephalothin, phenylbutazone,diphenylhydantoin, digitoxin and phenothiazine tranquilizers, andheparin.

In various embodiments, the patient treatment regimens include regimensin which one or more agents with neutropenic side effects areadministered. In exemplary embodiments, the one or more agent with aneutropenic side effect is selected from: cyclophosphamide, psychotropicdrugs and anticonvulsants such as clozapine and olanzapine, thionamides,ticlopidine, carbimazole, dapsone, dipyrone, methimazole, penicillin G,procainamide, propylthiouracil, trimethoprim, chloramphenicol,penicillins, cephalosporins, aminoglycosides, tetracyclines,nitroimidazoles, nitrofurantoin, flucytosine, rifampin, isoniazid,ethambutol, dapsone, sulfonamide antibiotics, clomiprimine,thiacetazone, dipyrone, sulfasalazine, mesalazine, ciprofloxacin,chloroquin, mebendazole, terbendafine, pyrimethamine, levamisole,ristocetin, griseofulvin, phenothiazines, benzodiazepines, amoxapine,meprobamate, barbiturates, risperidone, imipramine, desipramine,thiothixene, haloperidol, valproic acid, hydantoins, succinimides,trimethadione, carbamazepine, procainamide, quinidine, propafenone,captopril, propranolol, hydralazine, methyldopa, ibuprofen,indomethacin, sulindac, tolmetin, aspirin, aminopyine, phenylbutazone,diflunisal, benoxaprofen, allopurinol, colchicine, propylthiouracil,thiouracil, methimazole, carbimazole, thiocyanate, potassiumperchlorate, cimetidine, ranatadine, tripelennamine, methaphenilene,thenalidine, mianserin, bromopheneramine, quinine, hydroxychloroquin,quinacrine, diazoxide, dihydropyridines, vesnarinone, aprindine,imipenem/cilastatin, zidovudine, fludarabine, acyclovir, turbinafine,aminoglutethimide, famotidine, bezafibrate, flutamide, tamoxafen,penicillamine, retinoic acid, metoclopramide, phenindone, dinitrophenol,ethacrynic acid, rauwolfia, ethanol, chlorpropamide, tolbutamide,thiazides, spironolactone, methazolamide, acetazolamide, levodopa andcombinations thereof. See, Oyesanme et al., 1999, Psychosomatics, 40:5at p.414 421; the disclosure of which is incorporated herein byreference.

In certain embodiments, the myelosuppressive patient treatment regimencomprises one or more regimens suitable for the treatment of subjectsdiagnosed with acute myelogenous or myeloid leukemia (“AML”). Treatmentregimens for AML, typically consist of two phases, an initial phaseintended to induce remission, referred to as the induction phase, and asecond phase intended to prevent recurrence or relapse, referred to asthe consolidation phase. Treatments administered during the inductionphase are referred to as induction treatment regimens and treatmentsadministered during the consolidation phase are referred to asconsolidation treatment regimens. Standard induction treatment regimensinclude chemotherapy, referred to as induction chemotherapy, and areknown in the art. In an exemplary embodiment of induction chemotherapy,the chemotherapy regimen consists of treatment with cytarabine (araC)administered intravenously for 7 consecutive days and an anthracyclineagent (e.g., daunorubicin or idarubicin) administered on 3 consecutivedays. See Tallman, 2005, Hematology 2005:143-150; Robak et al., 2009,Clin. Therap. 31:2349-70. Consolidation treatment regimens can comprisechemotherapy, immunotherapy, bone marrow transplant, or combinationsthereof. In some embodiments, consolidation chemotherapy consists of oneor more cycles of the same chemotherapy regimen used during theinduction phase. In other embodiments, consolidation chemotherapyconsists of one or more cycles of high dose chemotherapy. Exemplaryembodiments of consolidation chemotherapy include two, three, four,five, or more cycles of treatment with cytarabine and an anthracyclineregimen as described above. In some embodiments, the consolidationchemotherapeutic regimen comprises a higher dose of the chemotherapeuticagent or agents administered during the induction phase. Theconsolidation phase can also comprise immunotherapy with one or moreagents such as, but not limited to, histamine dihydrochloride andinterleukin-2. In certain embodiments, the consolidation treatmentregimen comprises an allogeneic stem cell transplant. In variousembodiments, a PF4-interacting heparinoid is administered adjunctivelyto an induction and/or consolidation treatment regimen. In an exemplaryembodiment, ODSH is administered adjunctively to an induction and/orconsolidation treatment regimen.

5.1.2. Treatment Subjects

The subject to be treated (used interchangeably herein with “patient”)may be any animal, for example a mammal, preferably a human. In certainembodiments, the subject is an adult. In certain embodiments, thesubject is a child, for example a child diagnosed with a pediatriccancer.

In some embodiments, suitable subjects are patients diagnosed withcancer, and in need of an antineoplastic or cytotoxic treatment regimen.The cancer can be a solid tumor cancer in any organ or tissue, includingpancreatic cancer, ovarian cancer, uterine cancer, breast cancer,including metastatic breast cancer and chemotherapy-resistant breastcancer (e.g., breast cancer that recurs as a relapse within 6 months ofadjuvant chemotherapy with or without an anthracycline), head and neckcancer, bladder cancer, urothelial cancer, lung cancer (includingnon-small cell lung cancer), colorectal cancer, gastric cancer,esophageal cancer, neuroblastoma, liver cancer, melanoma, prostatecancer, osteosarcoma, and can be a hematologic cancer, such as lymphoma(including recurrent, Hodgkin's, and non-Hodgkin's lymphomas), andleukemia (including acute myelogenous leukemia, or AML, and pediatricacute lymphoblastic leukemia).

The methods described herein are particularly useful for cancers inwhich PF4 levels are elevated either in platelets or in the blood. Thus,in some embodiments, the subject has been diagnosed with a cancer inwhich PF4 levels are elevated either in platelets or in the blood. Incertain embodiments, the cancer is pancreatic cancer, colorectal cancer,osteosarcoma or leukemia (including acute myelogenous leukemia andpediatric acute lymphoblastic leukemia).

Suitable subjects for treatment also include subjects suffering from adisease or condition for which the recommended treatment regimen has amyelosuppressive side effect, including any of the treatment regimensdescribed above in Section 5.1.1.

In various embodiments, the suitable subject is a subject having anelevated level of PF4 in blood or in platelets, including variouscancers above-described and non-cancerous conditions with elevatedlevels of PF4 in blood or in platelets, a subject with an autoimmunedisease that can be treated with a treatment regimen including one ormore agents having a myelosuppressive side effect, such as rheumatoidarthritis, systemic lupus erythematosus, multiple sclerosis, Crohn'sdisease, ulcerative colitis, inflammatory bowel disease, or a subjecthaving decreased thrombopoietin levels, such as patients with livercancer, viral hepatitis, cirrhosis, or impaired liver function.

In various embodiments, the suitable subject is a subject who does nothave immune-mediated thrombocytopenia, thrombocytopenia due to anautoimmune condition, thrombocytopenia caused by increased destructionof platelets, or heparin-induced thrombocytopenia.

In embodiments in which the PF4-interacting heparin is partiallydesulfated, such as 2-O, 3-O desulfated heparin, there is a reduced riskof heparin-induced thrombocytopenia, even when administered incombination with heparin (unfractionated heparin or low molecular weightheparin) as an anticoagulant agent. See U.S. Pat. No. 7,468,358.Consequently, in some embodiments, the patient may be a subject who hasantibodies against heparin-PF4 complex and is at risk of heparin-inducedthrombocytopenia.

5.1.3. Other Adjunctive Agents and Therapy

The PF4-interacting heparinoid can be administered either as a soleagent adjunctive to a patient treatment regimen having amyelosuppressive side effect, or in combination with one or moreadditional agents or therapies.

Thus, in various embodiments, the methods further comprise adjunctiveadministration of one or more additional agents or therapies that arealso capable of attenuating thrombocytopenia and/or promotingthrombopoiesis, attenuating neutropenia and/or promoting granulopoiesis.In certain embodiments, the methods further comprise adjunctiveadministration of an anti-coagulating heparinoid. In some embodiments,two or more such agents and/or therapies are administered. The two ormore such agents can have the same activity (e.g.,anti-thrombocytopenic), different activity (e.g., a first agent ispro-thrombopoietic and a second agent is anti-neutropenic), oroverlapping activity (e.g., a first agent is pro-granulopoietic andanticoagulant and a second agent is anti-coagulant).

Suitable additional therapies or agents to attenuate thrombocytopeniaand/or promote thrombopoiesis include agents or therapies that act toincrease platelet count. Thus, in some embodiments, the one or moreadditional agent or therapy is selected from platelet transfusion,splenectomy, corticosteroids (e.g., prednisone and dexamethasone),platelet clearance inhibitors (e.g., danazol), thrombopoeitin,thrombopoietin mimetics (e.g., Nplate®, eltrombopag (Promacta®)),thrombopoietin receptor agonists (e.g., romiplostim and eltrombopag),interleukins, e.g. recombinant human interleukins (includinginterleukin-1, interleukin-3, interleukin-6, interleukin-11 (e.g.,Numega®)), lithium carbonate, and folate.

Suitable additional therapies or agents to attenuate neutropenia and/orpromote granulopoiesis include agents that act to increase neutrophilcount. Thus, in some embodiments, the one or more additional agent ortherapy is selected from recombinant human granulocyte colonystimulating factor (“G-CSF”) (filgrastim (Neupogen), pegfilgrastim(Neulasta)), and recombinant human granulocyte-macrophage colonystimulating factor (“GM-C SF”) (sargramostim (Leukine)).

In some clinical presentations, the patient may benefit fromanti-coagulation therapy. Thus, in some embodiments, the methodscomprise administering PF4-interacting heparinoid in combination withone or more anti-coagulating agents, such as one or more anti-coagulantheparinoids, preferably in such amounts, or in such ratios, as toprovide anticoagulation without risk of inducing or triggeringheparin-induced thrombocytopenia. In exemplary embodiments,anti-coagulation agents are selected from heparins, such asunfractionated heparin, and low molecular weight heparins, such asdalteparin, enoxaparin, fondaparinux, reviparin, and tinzaperin.Generally, the PF4-interacting heparinoid and the anti-coagulant areadministered in ratios in which the molar or weight amount ofPF4-interacting heparinoid exceeds the molar or weight amount ofanti-coagulant. In some embodiments, molar ratios of PF4-interactingheparinoid to anticoagulant heparin range from about 1:2 to about 10:1.Equivalent weight ratios are also contemplated. In some embodiments,weight ratios of PF4-interacting heparinoid to anticoagulant heparinrange about 1:1 to about 4:1.

In various embodiments, the one or more additional adjunctive agents ortherapies is administered concurrently, sequentially, or separately withPF4-interacting heparinoid. In some embodiments, the one or moreadditional agents or therapies is administered both concurrently andsequentially with PF4-interacting heparinoid.

5.2. Method of Promoting Thrombopoiesis

It has now been discovered that PF4-interacting heparinoids can increaseplatelet counts in human patients. As described in Example 3 below,patients treated with ODSH had increased platelet counts at the end of afirst 4 week cycle of a chemotherapy regimen for pancreatic cancer thathas a substantial myelosuppressive side effect. This effect continuedthrough successive cycles, with patients showing platelet counts abovelevels seen at screening (i.e., prior to treatment with ODSH), aftertwo, three, or even four cycles of adjunctive administration of ODSH andthe chemotherapy treatment regimen. Thus, in another aspect, methods forpromoting thrombopoiesis in a subject are provided. The methods compriseadministering an effective amount of a PF4-interacting heparinoid to thesubject. PF4-interacting heparinoids for use in the methods aredescribed below in Section 5.6. In an exemplary embodiment, thePF4-interacting heparinoid is ODSH. Suitable modes of administration anddosing regimens are described further below, in Section 5.8. Effectivedosages, and therapeutically effective amounts, of PF4-interactingheparinoid are described further below, in Section 5.9.

The method can be carried out in a thrombocytopenic subject or anon-thrombocytopenic subject.

In embodiments in which the subject is thrombocytopenic, thethrombocytopenia can be of varying etiology. Thus, in variousembodiments, the thrombocytopenia is (1) thrombocytopenia caused by atreatment regimen with a myelosuppressive side effect, as describedabove in Section 5.1.1, and the subjects may include those describedabove in Section 5.1.2, (2) thrombocytopenia caused by impairedproduction of platelets by the bone marrow, (3) thrombocytopenia causedby platelet sequestration in the spleen (splenomegaly), or (4)thrombocytopenia caused by increased destruction of platelets in theperipheral circulation, optionally due to an autoimmune condition.

In various embodiments, the subject's platelet count is reduced as aresult of—and, optionally, thrombocytopenia caused by—a disease orcondition. Accordingly, in certain embodiments, the subject is sufferingfrom an infection. In some embodiments, the infection results in sepsis,with or without disseminated intravascular coagulation. In someembodiments, subjects have elevated plasma levels of PF4, for example,more than about 5 ng/ml, more than about 6 ng/ml, more than about 7ng/ml, more than about 8 ng/ml, more than about 9 ng/ml, more than about10 ng/ml, more than about 11 ng/ml, more than about 12 ng/ml, more thanabout 15 ng/ml, more than about 17 ng/ml, more than about 20 ng/ml, morethan about 22 ng/ml, more than about 25 ng/ml, more than about 27 ng/ml,more than about 30 ng/ml, more than about 40 ng/ml, up to about 45ng/ml, up to about 50 ng/ml or greater. Lorenz et al., 1988, Infection16(5):273-6 and PF4 assay therein. In some embodiments, the subjectshave thrombocytopenia that is not heparin-induced thrombocytopenia.

In exemplary embodiments, the thrombocytopenia is selected fromradiation-induced thrombocytopenia; drug-induced thrombocytopenia;consumption thrombocytopenia; immune-mediated thrombocytopenia,including alloimmune thrombocytopenia and auto-immune thrombocytopenia,including immune thrombocytopenic purpura (or ITP); infectious cyclicthrombocytopenia; myelophthisic thrombocytopenia caused by neoplasticinvasion of the bone marrow; surface-induced thrombocytopenia;vaccine-induced thrombocytopenia; liver, bone marrow or stem celltransplant-induced thrombocytopenia; and thrombocytopenia attendant toautoimmune disease (e.g., rheumatoid arthritis, systemic lupuserythematosus) or lymphoproliferative disorder (e.g., chroniclymphocytic leukemia). In some embodiments, the thrombocytopenia is notor is other than: immune-mediated thrombocytopenia, thrombocytopenia dueto an autoimmune condition or thrombocytopenia caused by increaseddestruction of platelets. In some embodiments, the thrombocytopenia isother than heparin-induced thrombocytopenia.

In various embodiments, the methods of promoting thrombopoiesis areuseful for treating a subject who has radiation-induced thrombocytopeniacaused by radiation therapy or by non-therapeutic exposure to ionizingradiation, for example a subject who has radiation poisoning orradiation sickness as a result of a radiological or nuclear accident orattack.

In various embodiments, suitable subjects include patients who wouldbenefit from an increased platelet count, such as in advance of surgery,transfusion, therapy with a treatment regimen having a myelosuppressiveside effect, or other procedure or treatment that could lower plateletcount or increase the need for clotting. In exemplary embodiments, thesubject is diagnosed with cancer, and/or is in need of surgery,transfusion, or therapy with a treatment regimen having amyelosuppressive side effect. In some embodiments, the subject is atrisk for radiation-induced thrombocytopenia due to radiation therapy ornon-therapeutic exposure to ionizing radiation, for example as a resultof a radiological or nuclear accident or attack.

5.2.1. Additional Agents and Therapy

In methods of promoting thrombopoiesis, the PF4-interacting heparinoidcan be administered either as a sole agent or in adjunctive combinationwith one or more additional agents or therapies. In some embodiments,the one or more additional agents or therapy is capable of promotingthrombopoiesis. In various embodiments, the one or more additionalagents or therapy is anti-coagulating.

Suitable additional therapies or agents to promote thrombopoiesisinclude agents or therapies that act to increase platelet count. Inexemplary embodiments, the one or more additional agents or therapy isselected from platelet transfusion, splenectomy, corticosteroids (e.g.,prednisone and dexamethasone), platelet clearance inhibitors (e.g.,danazol), thrombopoeitin, thrombopoietin mimics, thrombopoietin receptoragonists (e.g., romiplostim and eltrombopag), interleukins, e.g.recombinant human interleukins (including interleukin-1, interleukin-3,interleukin-6, interleukin-11), lithium carbonate, and folate. Otheradditional agents and therapy are described in Section 5.1.3.

In some clinical presentations, the patient may need anti-coagulationtherapy, despite requiring thrombopoiesis. Thus, in some embodiments,the PF4-interacting heparinoid is administered adjunctive to one or moreanti-coagulating agents, preferably without risk of inducing ortriggering heparin-induced thrombocytopenia. Anti-coagulation agentsinclude heparins, such as unfractionated heparin, and low molecularweight heparins, such as dalteparin, enoxaparin, fondaparinux,reviparin, tinzaperin. Generally, the PF4-interacting heparinoid and theanti-coagulant can be administered in ratios in which the amount ofPF4-interacting heparinoid exceeds the amount of anti-coagulant. Molarratios of PF4-interacting heparinoid to anticoagulant heparin range fromabout 1:2 to about 10:1. Equivalent weight ratios are also contemplated.

5.3. Method of Promoting Neutrophil Production

It has now been discovered that PF4-interacting heparinoids can increaseneutrophil counts in human patients. As further described in Example 3below, patients treated with ODSH had increased neutrophil counts at theend of a first 4 week cycle of chemotherapy, despite receivingconcurrent treatment with a chemotherapeutic regimen havingmyelosuppressive side effects, and consistently showed increasedneutrophil counts at the end of successive 4-week cycles relative toneutrophil counts mid-cycle. In some instances, patients showedneutrophil counts above levels seen at screening (i.e., prior totreatment with ODSH), after two, three, or even four cycles ofadjunctive administration of ODSH and the chemotherapy treatmentregimen. Thus, in another aspect, methods for promoting neutrophilproduction in a subject are presented. The methods compriseadministering an effective amount of a PF4-interacting heparinoid, asdescribed further in Section 5.6 below, to the subject. In an exemplaryembodiment, the PF4-interacting heparinoid is ODSH. Suitable modes ofadministration and dosing regimens are described further below, inSection 5.8. Effective dosages, and therapeutically effective amounts,of PF4-interacting heparinoid are described further below, in Section5.9.

The method can be carried out in a neutropenic subject or anon-neutropenic subject. The subject can be an adult or a child. Invarious embodiments, the subjects include those described above atSection 5.1.2.

In embodiments in which the subject to be treated is neutropenic, theneutropenia may be chronic or acute. In various embodiments, theneutropenia is congenital (e.g., caused by Kostmann's Syndrome),cyclical or idiopathic. In some embodiments, the neutropenia issecondary to another condition, such as cancer, viral infection (e.g.,acquired immunodeficiency syndrome (AIDS)). In some embodiments, theneutropenia is autoimmune. In some embodiments, neutropenia is caused byinfiltration and destruction of bone marrow due to leukemia, myeloma,lymphoma or a metastatic solid tumor such as, for example, breast orprostate cancer. In some embodiments, the neutropenia isradiation-induced neutropenia, resulting from intentional ornon-therapeutic exposure to ionizing radiation, for example as a resultof a radiological or nuclear accident or attack.

Neutropenia can be a side effect of agents or procedures. Thus, in someembodiments, neutropenia is caused by a treatment regimen having amyelosuppressive side effect, e.g., chemotherapy, radiation therapy forcancer, bone marrow transplantation associated with cancer therapy andas described above in Section 5.1.1.

In some embodiments, neutropenia is immune-mediated, includingautoimmune or alloimmune (e.g., caused by a non-self antigen thatstimulates antibody formation and causes a hypersensitive reaction).

In various embodiments, the methods of promoting neutrophil productionare useful for treating a subject diagnosed with radiation-inducedneutropenia caused by radiation therapy or by non-therapeutic exposureto ionizing radiation, for example as a result of a radiological ornuclear accident or attack.

The methods of promoting neutrophil production are also useful fortreating non-neutropenic subjects. Suitable subjects include thosedescribed above in Section 5.1.2. Such methods are particularly usefulwhere the subject would benefit from an increased neutrophil count. Inexemplary embodiments, the subject has been diagnosed with cancer,and/or is in need of surgery, or therapy with a treatment regimen havinga myelosuppressive side effect. In some embodiments, the subject is atrisk for radiation-induced neutropenia due to radiation therapy ornon-therapeutic exposure to ionizing radiation, for example as a resultof a radiological or nuclear accident or attack. In some embodiments,the subject is at increased risk of contracting an infection.

5.3.1. Additional Agents and Therapy

In the methods of promoting neutrophil production, the PF4-interactingheparinoid can be administered either as a sole agent or in adjunctivecombination with one or more additional agents or therapies. In someembodiments, the one or more additional agents or therapy are capable ofpromoting neutrophil production. In various embodiments, the one or moreadditional agents or therapies are anti-coagulating.

Suitable therapies or agents to promote neutrophil production includeagents that act to increase absolute neutrophil count. In exemplaryembodiments, the one or more additional agent is selected fromrecombinant human granulocyte colony stimulating factor (“G-CSF”)(filgrastim (Neupogen), pegfilgrastim (Neulasta)) and recombinant humangranulocyte-macrophage colony stimulating factor (“GM-CSF”)(sargramostim(Leukine)).

In some clinical situations, the patient may need anti-coagulationtherapy. Thus, in some embodiments, the PF4-interacting heparinoid isadministered adjunctive to one or more anti-coagulating agents,preferably without inducing or triggering heparin-inducedthrombocytopenia. Anti-coagulation agents for use in such embodimentsinclude, for example, heparins, such as unfractionated heparin, and lowmolecular weight heparin, such as dalteparin, enoxaparin, fondaparinux,reviparin, tinzaperin. In various embodiments, the PF4-interactingheparinoid and the anti-coagulant are administered in ratios in whichthe amount of PF4-interacting heparinoid exceeds the amount ofanti-coagulant. Molar ratios of PF4-interacting heparinoid toanticoagulant heparin range from about 1:2 to about 10:1. Equivalentweight ratios are also contemplated.

Other suitable additional agents and therapies are described in Section5.1.3 above.

5.4. Method of Increasing Efficacy of Treatment Regimens withMyelosuppressive Side Effects

Myelosuppressive side effects such as thrombocytopenia and neutropeniathat are caused by patient treatment regimens can be dose-limiting,limiting either the dose amount, the frequency of administration, orboth, thereby decreasing the efficacy of the patient treatment regimen.Attenuating such myelosuppressive side effects would permit the doseamount and/or frequency of treatment to be maintained or increased,which should in turn lead to greater efficacy of the patient treatmentregimen. As shown in Example 3 below, ODSH, a PF4-interactingheparinoid, attenuated the myelosuppressive side effects caused by anantineoplastic treatment regimen for pancreatic cancer; in particular,administering ODSH adjunctively with the chemotherapy treatmentincreased both platelet and neutrophil counts above those beforetreatment. Thus, administration of a PF4-interacting heparinoidadjunctively with such a treatment regimen should permit the dose amountand/or frequency of treatment to be maintained or increased, therebyincreasing efficacy of the patient treatment regimen.

Consequently, in another aspect, methods for increasing efficacy of atreatment regimen with a myelosuppressive effect are provided. Themethods comprise administering a therapeutically effective amount of aPF4-interacting heparinoid to the subject patient as an adjunct to thepatient treatment regimen having myelosuppressive side effect, such asthe ICE regimen, without reducing the dose and/or dosage frequency ofthe myelosuppressive patient treatment following a reference treatmentadministration or treatment cycle. In some embodiments, the referencetreatment administration or cycle is the first treatment administrationor treatment cycle. In various embodiments, the reference treatmentadministration or treatment cycle of the patient treatment regimen issubsequent to the first treatment administration or treatment cycle.

In some embodiments, the method further comprises administering a dosehigher than is typically used for such administration or cycle in theabsence of adjunctive administration of a PF4-interacting heparinoid.

In some embodiments, the methods further comprise determining an initialplatelet and/or neutrophil count in a sample of blood from a patient.

In various embodiments, the PF4-interacting heparinoid is administeredin an amount that is effective to raise the patient's platelet and/orneutrophil count above a prior-determined threshold level. In certainembodiments, the prior-determined threshold level is the level belowwhich administration of the patient treatment regimen having amyelosuppressive side effect is contraindicated. Suitable modes ofadministration and dosing regimens are described further below, inSection 5.8. Effective dosages, and therapeutically effective amounts,of PF4-interacting heparinoid are described further below, in Section5.9.

In certain embodiments, the methods comprise determining an initialplatelet count in a sample from a patient, and then administering anamount of a PF4-interacting heparinoid effective to raise the patient'splatelet count above a threshold level below which therapy with apatient treatment regimen having a myelosuppressive side effect iscontraindicated. In various embodiments, an amount of a PF4-interactingheparinoid is administered sufficient to maintain platelet levels abovelevels that indicate grade 4 or grade 3 thrombocytopenia. In variousembodiments, an amount of a PF4-interacting heparinoid is administeredsufficient to maintain platelet levels above levels that indicate grade2 or grade 1 thrombocytopenia. Optionally, the methods can furthercomprise administering adjunctively to the PF4-interacting heparinoidone or more agents or therapies that is anti-thrombocytopenic,anti-neutropenic, anticoagulant, or has some other therapeutic activity.In some embodiments, the methods comprise a further step ofadministering a patient treatment regimen having a myelosuppressive sideeffect to the patient whose platelet count is above a level thatcontraindicates such therapy. Optionally, the dose amount and/orfrequency of the patient treatment regimen can be increased.

In certain embodiments, the methods comprise determining an initialneutrophil count in a blood sample from a patient and administering anamount of a PF4-interacting heparinoid effective to raise the patient'sneutrophil count above a threshold level below which therapy withpatient treatment regimen having a myelosuppressive side effect iscontraindicated. In various embodiments, an amount of a PF4-interactingheparinoid is administered sufficient to maintain neutrophil levelsabove levels that indicate of grade 4 or grade 3 neutropenia, i.e.,above about 1000 neutrophils/μl of blood and above about 500neutrophils/μl of blood. In various embodiments, an amount of aPF4-interacting heparinoid is administered sufficient to maintainneutrophil levels above levels that indicate of grade 2 or grade 1neutropenia. Optionally, the methods can further comprise administeringadjunctive to the PF4-interacting heparinoid one or more agents ortherapies that is anti-neutropenic, anti-thrombocytopenic,anticoagulant, or has some other therapeutic activity. In someembodiments, the methods comprise a further step of administering apatient treatment regimen having a myelosuppressive side effect to thepatient whose neutrophil count is above a level that contraindicatessuch therapy. Optionally, the dose amount and/or frequency of thepatient treatment regimen can be increased.

5.5. Method of Enhancing Efficacy of Antineoplastic Patient TreatmentRegimens

It has been discovered that adjunctive administration of ODSH, aPF4-interacting heparinoid, enhances the ability of antineoplastictreatment regimens to inhibit tumor growth. As shown in Examples 1 and 2below, adjunctive administration of ODSH results in greater inhibitionof tumor growth in murine xenograft models of pancreatic and ovariancancer, than administration of either ODSH or a chemotherapeutictreatment regimen alone. Thus, in another aspect, methods are providedherein for enhancing the efficacy of an antineoplastic treatmentregimen. The methods comprise administering a therapeutically effectiveamount of a PF4-interacting heparinoid to a subject patient as anadjunct to an antineoplastic treatment regimen.

Antineoplastic treatment regimens are patient treatment regimens usefulin treating cancer. Suitable antineoplastic treatment regimens includechemotherapeutic treatment regimens (including induction chemotherapyand consolidation chemotherapy), antibody treatment regimens, andcombinations thereof, as described further in Section 5.1.1 above.

The subject to be treated may be any animal, for example a mammal,preferably a human. In certain embodiments, the subject is an adult. Incertain embodiments, the subject is a child. Subjects to be treated arepatients in need of antineoplastic treatment, in particular patientssuffering from, or diagnosed with, cancer. The cancer can be in anyorgan or tissue, including pancreatic cancer, ovarian cancer, uterinecancer, breast cancer, including metastatic breast cancer andchemotherapy-resistant breast cancer (e.g., breast cancer that recurs asa relapse within 6 months of adjuvant chemotherapy with or without ananthracycline), head and neck cancer, bladder cancer, urothelial cancer,lung cancer (including non-small cell lung cancer), colorectal cancer,gastric cancer, esophageal cancer, lymphoma (including recurrent,Hodgkin's, and non-Hodgkin's lymphomas), liver cancer, melanoma,prostate cancer, osteosarcoma and leukemia (including acute myelogenousleukemia and pediatric acute lymphoblastic leukemia).

The methods described herein are particularly useful for cancers inwhich PF4 levels are elevated either in platelets or in the blood. Thus,in some embodiments, the subject has been diagnosed with a cancer inwhich PF4 levels are elevated either in platelets or in the blood. Incertain embodiments, the cancer is pancreatic cancer, colorectal cancer,osteosarcoma or leukemia (including acute myelogenous leukemia andpediatric acute lymphoblastic leukemia).

PF4-interacting heparinoids suitable for use in the methods aredescribed below in Section 5.6. In an exemplary embodiment, thePF4-interacting heparinoid is ODSH. Suitable modes of administration anddosing regimens are described further below, in Section 5.8. Effectivedosages and therapeutically effective amounts of PF4-interactingheparinoid are described further below, in Section 5.9.

5.6. PF4-Interacting Heparinoids

The heparinoids for use in the methods described herein are heparinoidsthat are capable of interacting with PF4, and counteracting PF4′sability to suppress production of platelets and neutrophils. As usedherein, PF4-interacting heparinoids include heparinoids which bind PF4and heparinoids which compete with PF4 for binding to progenitor cellsin the myeloid cell lineage, e.g., megakaryocytes. A specific assay forbinding of a heparinoid to PF4 is provided in Joglekar et al., 2012,Thromb Haemost 107(4):717-725, the disclosure of which is incorporatedby reference herein. In some embodiments, a PF4-interacting heparinoidis a heparinoid that competes for binding to PF4 with unfractionatedheparin, as determined by a competition assay, see e.g., Stringer etal., 1997, J. Biol. Chem. 272(33) 20508-20514, the disclosure of whichis incorporated by reference herein.

PF4-interacting heparinoids are linear glycosaminoglycan polymers madeup of alternating or repeating iduronic acid and glucosamine unitsbearing O-sulfate, N-sulfate, and N-acetyl substitutions. Preferably,PF4-interacting heparinoids for use in the methods described herein arepolymers having an average molecular weight of at least about 8 kDa, forexample having an average molecular weight ranging from about 8 kDa toabout 15 kDa. In certain embodiments, the PF4-interacting heparinoidshave an average molecular weight of greater than about 8 kDa. Morepreferably, PF4-interacting heparinoids for use in the methods describedherein have an average molecular weight that ranges in size from about11 kDa to about 13 kDa. Molecular weight of heparinoids can bedetermined by high performance size exclusion chromatography as is knownin the art. See, e.g., Lapierre et al., 1996, Glycobiology 6(3):355-366,at page 363; Fryer et al., 1997, J. Pharmacol. Exp. Ther. 282: 208-219,at page 209.

Optionally, the PF4-interacting heparinoid does not cause plateletactivation and heparin-induced thrombocytopenia (HIT), and is thereforeuseful for treating subjects at risk of heparin-inducedthrombocytopenia, including subjects with antibodies against aPF4/heparin complex. Thus, in various embodiments, the PF4-interactingheparinoid does not trigger platelet activation that leads toheparin-induced thrombocytopenia; platelet activation can be determinedusing a serotonin release assay, as described in U.S. Pat. No. 7,468,358and Sheridan et al., 1986, Blood 67:27-30. In some embodiments, thePF4-interacting heparinoid binds PF4 but is not recognized byanti-heparin-PF4 complex antibodies, even when complexed with PF4.

In various preferred embodiments, the PF4-interacting heparinoid issubstantially nonanticoagulant. Anti-coagulation activity can bedetermined using assays known in the art, e.g., activated partialthromboplastin time (aPTT), prothrombin time, anti-X_(a) clotting andamidolytic assays. See, e.g., U.S. Pat. No. 5,668,118, Example IV; Fryeret al., 1997, J. Pharmacol. Exp. Ther. 282: 208-219, at page 209; Rao etal., 2010, Am. J. Physiol. 299:C97-C110, at page C98; United StatesPharmacopeial Convention 1995 (for USP anticoagulant assay andamidolytic assay).

In typical embodiments, the PF4-interacting heparinoids are partiallydesulfated. Preferably, the PF4-interacting heparinoids aresubstantially desulfated at the 2-O position of α-L-iduronic acid(referred to herein as the “2-O position”) and/or desulfated at the 3-Oposition of D-glucosamine-N-sulfate (6-sulfate) (referred to herein asthe “3-O position”). In some embodiments, the PF4-interactingheparinoids are at least 85%, at least 90%, at least 95%, or at least99% desulfated at the 2-O position. In some preferred embodiments, thePF4-interacting heparinoids are at least 99% desulfated at the 2-Oposition. In some embodiments, the PF4-interacting heparinoids are atleast 85%, at least 90%, at least 95%, or at least 99% desulfated at the3-O position. In some preferred embodiments, the PF4-interactingheparinoids are at least 99% desulfated at the 3-O position. In someembodiments, the PF4-interacting heparinoids are at least 85%, at least90%, at least 95%, at least 99% desulfated at the 2-O position and the3-O position. In some preferred embodiments, the PF4-interactingheparinoids are at least 99% desulfated at the 2-O position and the 3-Oposition.

In typical embodiments, the PF4-interacting heparinoid comprisessubstantially N-sulfated and 6-O sulfated D-glucosamine. In someembodiments, the carboxylates on α-L-iduronic acid sugars ofPF4-interacting heparinoid are substantially intact.

An exemplary PF4-interacting heparinoid is substantially 2-O, 3-Odesulfated heparin, referred to herein as ODSH. ODSH for use in theabove-described methods can be prepared from bovine or porcine heparin.In an exemplary method of preparing ODSH from porcine heparin, ODSH issynthesized by cold alkaline hydrolysis of USP porcine intestinalheparin, which removes the 2-O and 3-O sulfates, leaving N- and 6-Osulfates on D-glucosamine sugars and carboxylates on α-L-iduronic acidsugars substantially intact. Fryer, A. et al., 1997, J. Pharmacol. Exp.Ther. 282: 208-219. Using this method, ODSH can be produced with anaverage molecular weight of about 11.7±0.3 kDa.

In contrast to unfractionated heparin, ODSH is substantiallynon-anticoagulating: administered to a subject at a dose that isequivalent to a fully-anticoagulating dose of unfractionated heparin,the clotting time measured in an aPTT assay is no greater than 45seconds, and typically in the upper range of normal, where normalclotting time ranges from about 27 to 35 seconds. By comparison,unfractionated heparin administered to a subject at a fullyanticoagulant dose causes time to clot to range from about 60 to about85 seconds in an aPTT assay. Another measure of ODSH's anticoagulantactivity is its anti-X_(a) activity which can be determined in an assaycarried out using plasma treated with Russell viper venom. In specificexamples, ODSH exhibited less than 9 U of anticoagulant activity/mg inthe USP anticoagulant assay (e.g., 7±0.3 U), less than 5 U of anti-X_(a)activity/mg (e.g., 1.9±0.1 U/mg) and less than 2 U of anti-II_(a)activity/mg (e.g., 1.2±0.1 U/mg) (compare to unfractionated heparinwhich has an activity of 165-190 U/mg in all three assays). See Rao etal., 2010, Am. J. Physiol. 299:C97-C110, page C101. Whether or not aheparinoid is substantially non-anticoagulating can be determined usingany of the above assays. Furthermore, ODSH has a low affinity foranti-thrombin III (Kd˜339 μM or 4 mg/ml vs. 1.56 μM or 22 μg/ml forunfractionated heparin), consistent with the observed low level ofanticoagulant activity, measured as described in Rao et al., supra, atpage C98.

Methods for the preparation of 2-O, 3-O desulfated heparin may also befound, for example, in U.S. Pat. Nos. 5,668,118, 5,912,237, and6,489,311, and WO 2009/015183, the contents of which are incorporatedherein in their entirety, and in U.S. Pat. Nos. 5,296,471, 5,969,100,and 5,808,021.

5.7. Pharmaceutical Compositions and Unit Dosage Forms

In typical embodiments, the PF4-interacting heparinoid will beadministered in the form of a pharmaceutical formulation or composition.Pharmaceutical compositions, suitable for administration to subjects,may optionally include additional active and/or therapeutic agents, asis known in the art. See Remington: The Science and Practice ofPharmacy, 21^(st) Ed. (2005), Lippincott Williams & Wilkins,incorporated herein by reference. The formulations will typicallyinclude one or more pharmaceutically acceptable carriers, excipients, ordiluents. The specific carriers, excipients, and/or diluents used willdepend on the desired mode of administration.

In various embodiments, the pharmaceutical compositions is in the formof a sterile, non-pyrogenic, fluid composition.

The pharmaceutical compositions can be formulated for administration tosubjects by a variety of routes, including intranasally, by inhalation,intramuscularly, intraperitoneally, and parenterally, includingintravenously or subcutaneously. Pharmaceutical compositions can beformulated in volumes and concentrations suitable for bolusadministration, for continuous infusion, or for subcutaneousadministration.

Pharmaceutical compositions can be conveniently presented in unit dosageforms which contain a predetermined amount of PF4-interactingheparinoid. In various embodiments, unit dosage forms of PF4-interactingheparinoid contain 1 mg to 1 g, or 5 mg to 500 mg of PF4-interactingheparinoid.

5.8. Modes of Administration

PF4-interacting heparinoids can be administered in the methods describedherein by a variety of routes, as noted above. In presently preferredembodiments, the PF4-interacting heparinoid is administeredintravenously, either as one or more boluses, as a continuous infusion,or as one or more boluses followed by continuous infusion.

In a variety of embodiments, PF4-interacting heparinoid is administeredfor a period of 1 day to indefinitely, a period of 1 week to 6 months, aperiod of 3 months to 5 years, a period of 6 months to 1 or 2 years, orthe like. Optionally, PF4-interacting heparinoid administration isrepeated; for example, in certain embodiments, PF4-interactingheparinoid is administered once daily, twice daily, three times daily,four times daily, five times daily, every two days, every three days,every five days, once a week, once every two weeks, once a month, everyother month, semi-annually, or annually. In certain embodiments,PF4-interacting heparinoid is administered at regular intervals over aperiod of several weeks, followed by a period of rest, during which noPF4-interacting heparinoid is administered. For example, in certainembodiments, PF4-interacting heparinoid is administered for one, two,three, or more weeks, followed by one, two, three, or more weeks withoutPF4-interacting heparinoid administration. The repeated administrationcan be at the same dose or at a different dose. PF4-interactingheparinoid can be administered in one or more bolus injections, one ormore infusions, or one or more bolus injections followed or preceded byinfusion.

In embodiments in which PF4-interacting heparinoid is administeredadjunctively to a patient treatment regimen having a myelosuppressiveregimen, and/or adjunctively to administration of one or more additionalagent(s) or therap(ies) having anti-thrombocytopenic,pro-thrombopoietic, anti-neutropenic, and/or pro-granulopoieticactivity, the PF4-interacting heparinoid is administered intherapeutically effective temporal proximity to the treatment regimenhaving a myelosuppressive side effect and/or additional agent ortherapy. Administration of a PF4-interacting heparinoid can beconcurrent with (at the same time), sequential to (at a different timebut on the same day, e.g., during the same patient visit), or separatefrom (on a different day) administration of the patient treatmentregimen having a myelosuppressive side effect or other agent or therapy.In various embodiments, the adjunctively administered PF4-interactingheparinoid is administered concurrently, sequentially, and/or separatelyfrom the patient treatment regimen having myelosuppressive side effector other agent or therapy being administered. When administeredsequentially or separately, PF4-interacting heparinoid can beadministered before, after, or both before and after the othertreatment.

In embodiments in which PF4-interacting heparinoid is administeredadjunctively, the PF4-interacting heparinoid can be administrated viathe same or different route as the other treatment administered intemporal proximity. In various embodiments, PF4-interacting heparinoidis administered concurrently or sequentially by the same route. Forexample, in certain embodiments, PF4-interacting heparinoid and othertreatment are administered intravenously, either concurrently orsequentially. Optionally, as part of a treatment regimen, thePF4-interacting heparinoid can further be administered separately (on adifferent day) from the other treatment by a different route, e.g.,subcutaneously. In specific embodiments, PF4-interacting heparinoid isadministered intravenously on the same day, either at the same time(concurrently), a different time (sequentially), or both concurrentlyand sequentially with other treatment, and is also administeredsubcutaneously on one or more days when the patient is not receivingother treatment. In various embodiments, PF4-interacting heparinoid isadministered concurrently or sequentially by a different route.Optionally, as part of a treatment regimen, the PF4-interactingheparinoid can further be administered separately (on a different day)from the other treatment by the same or different route as that by whichthe other treatment is administered.

In one embodiment, PF4-interacting heparinoid is administered on days 1,8, and 15 of a 28-day chemotherapy cycle as an initial bolus followed bya 48-hour continuous infusion. The course of treatment can furtherinclude administration of one or more chemotherapeutic agentssequentially before or after PF4-interacting heparinoid. Optionally,PF4-interacting heparinoid is administered subcutaneously on day 21and/or one, several, or all of days 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13,14, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, and 27.

5.9. Effective Dosages

PF4-interacting heparinoid is administered to the subject in an amountsufficient or effective to provide a therapeutic benefit, i.e., atherapeutically effective amount. The therapeutically effective amountdepends on the therapeutic benefit that is sought—e.g., attenuation ofmyelosuppressive side effects such as thrombocytopenia and neutropenia,and/or promotion of thrombopoiesis, and/or promotion of granulopoiesis,and/or enhancement of antineoplastic effect.

In methods in which PF4-interacting heparinoid is administered toattenuate a myelosuppressive side effect of a patient treatment regimen,a myeloprotective amount of PF4-interacting heparinoid is administered,that is, an amount sufficient, in typical embodiments, to achieve one ormore of the following, as compared to historical data on the identicalpatient treatment regimen without adjunctive administration ofPF4-interacting heparinoid:

-   -   (a) thrombocytopenia is improved by at least one grade (e.g.,        from grade 4 to grade 3, 2, 1, or 0; from grade 3 to grade 2, 1,        or 0; from grade 2 to grade 1 or 0; or from grade 1 to grade 0);    -   (b) platelet count is increased by at least 10%, at least 20%,        at least 30%, at least 40%, at least 50%, at least 100%, at        least 200%;    -   (c) platelet count is increased by at least 5,000, at least        10,000, at least 15,000, at least 20,000, at least 25,000, at        least 30,000 platelets per μl of blood;    -   (d) neutropenia is improved by at least one grade (e.g., from        grade 4 to grade 3, 2, 1, or 0; from grade 3 to grade 2, 1, or        0; from grade 2 to grade 1 or 0; or from grade 1 to grade 0);    -   (f) absolute neutrophil count is increased by at least 10%, at        least 20%, at least 30%, at least 40%, at least 50%, at least        100%, at least 200%;    -   (d) absolute neutrophil count has increased by at least 500, at        least 1000, at least 1500, at least 2000, at least 2500, at        least 3000 neutrophils per μl of blood.

In particular embodiments, an amount of PF4-interacting heparinoid isadministered sufficient to achieve one or more of the above-describedeffects as compared to pre-treatment levels.

In methods in which PF4-interacting heparinoid is administered topromote thrombopoiesis, a thrombopoietically-effective amount of aPF4-interacting heparinoid is, in typical embodiments, an amounteffective to cause a measureable rise a subject's platelet count ascompared to pre-treatment levels.

In methods in which PF4-interacting heparinoid is administered topromote neutrophil production, a granulopoietically-effective amount ofa PF4-interacting heparinoid is, in typical embodiments, an amounteffective to cause a measureable rise a subject's absolute neutrophilcount as compared to pre-treatment levels.

In methods in which PF4-interacting heparinoid is administered toenhance efficacy of an antineoplastic treatment regimen, atherapeutically effective amount of a PF4-interacting heparinoid is anamount effective or sufficient to provide a therapeutic benefit. In thecontext of enhancing the efficacy of an antineoplastic treatmentregimen, in various embodiments, a therapeutic benefit is achievement ofone or more of the following: halting or slowing the growth of tumors,reducing the size and/or number of tumors within a patient, increasinglife expectancy, reduction in constitutional side effects of theantineoplastic treatment (e.g., weight loss, loss of appetite, nausea,vomiting, fatigue), permitting reduction in dosage or frequency ofdosage of the antineoplastic treatment regimen without reduced efficacy,and/or improving patient quality of life. A complete cure, whiledesirable, is not required for therapeutic benefit to exist. In somecontexts, a therapeutic benefit can be correlated with one or moresurrogate end points, in accordance with the knowledge of one ofordinary skill in the art. By way of example and not limitation,enhancing the efficacy of an antineoplastic treatment regimen can bemeasured in vivo. Exemplary in vivo assays for measuring tumor growthinhibition are described below for two different cancers and twodifferent antineoplastic treatment regimens in Examples 1 and 2 below.

The amount of PF4-interacting heparinoid administered will depend onvarious factors, including whether the subject is thrombocytopenicand/or neutropenic, the severity of any such thrombocytopenia and/orneutropenia, whether PF4-interacting heparinoid is being administeredadjunctively to a patient treatment regimen, and the age and conditionof the subject being treated, among others. The appropriate dosage canbe readily determined by a person of skill in the art. In practice, aphysician will determine appropriate dosages to be used. This dosage canbe repeated as often as appropriate. The amount and/or frequency of thedosage can be altered, increased, or reduced, depending on the subject'sresponse and in accordance with standard clinical practice. The properdosage and treatment regimen can be established by monitoring theprogress of therapy using conventional techniques known to skilledartisans.

In some embodiments, PF4-interacting heparinoid is administered at adose or amount per kilogram of patient body weight ranging from about 1mg/kg to about 25 mg/kg for intravenous bolus doses, and from about 0.1mg/kg/hr to about 2.5 mg/kg/hr for intravenous infusions. In a specificembodiment, PF4-interacting heparinoid is administered as an intravenousbolus at a dose of about 4 mg/kg, optionally followed by an intravenousinfusion of PF4-interacting heparinoid at a dose of about 0.375 mg/kg/hrfor 48 hours. In typical embodiments, a bolus dose is administered overless than a minute, about a minute, about 2 minutes, about 3 minutes,about 4 minutes, or about 5 minutes. For subcutaneous administration,PF4-interacting heparinoid can be administered at doses ranging fromabout 25 mg to about 400 mg, in volumes of 2.0 mL or less per injectionsite.

Pharmaceutical compositions of PF4-interacting heparinoid can beformulated in an amount that permits bolus intravenous administrationand/or continuous intravenous infusion at such doses. In one embodiment,the pharmaceutical composition comprises PF4-interacting heparinoid in asterile vial at a concentration of 50 mg/mL. When formulated forsubcutaneous administration, pharmaceutical compositions can containPF4-interacting heparinoid at a concentration ranging from 50 mg/ml to350 mg/ml suitable for administration at doses ranging from about 25 toabout 400 mg, in volumes of 2.0 mL or less per injection site.

6. EXAMPLES 6.1. Example 1 ODSH, a PF4-Interacting Heparinoid, Enhancesthe Efficacy of Gemcitabine in an In Vivo Murine Xenograft Model ofHuman Pancreatic Cancer and Demonstrates Antineoplastic Effect whenAdministered Alone

This experiment demonstrates that adjunctive administration of ODSHenhances the efficacy of gemcitabine against human pancreatic tumorsgrowing as xenografts in athymic nude mice, and demonstrates that ODSHinhibits tumor growth when administered alone.

Materials & methods. Compounds tested in the experiment were as follows.ODSH was made by Pyramid Laboratories, Inc. (Costa Mesa, Calif.). ODSHwas provided at a stock concentration of 50 mg/ml and stored at roomtemperature until use. ODSH was diluted in a 0.9% NaCl solution (B.Braun Medical Inc., Irvine, Calif.) to a concentration of 2.4 mg/ml todeliver 24 mg/kg, in a 10 ml/kg dose volume when administeredintravenously. A concentration of 4.8 mg/ml was formulated to deliver a24 mg/kg dose at a 5 ml/kg dose volume when administered subcutaneously.ODSH was formulated fresh prior to each dose.

The chemotherapeutic agents oxaliplatin, gemcitabine, and nab-paclitaxelwere also tested. Oxaliplatin was manufactured by Sanofi-Aventis(Bridgewater, N.J.) and diluted in a 0.9% NaCl solution to aconcentration of 1 mg/ml to deliver 10 mg/kg, in a 10 ml/kg dose volume.Gemcitabine was manufactured by Eli Lilly and Co. (Indianapolis, Ind.)and diluted in a 0.9% NaCl solution to a concentration of 8 mg/ml todeliver 80 mg/kg, in a 10 ml/kg dose volume. Nab-paclitaxel wasmanufactured by Abraxis BioScience LLC (Bridgewater, N.J.) and dilutedin a 0.9% NaCl solution to a concentration of 1.5 mg/ml to deliver 15mg/kg, in a 10 ml/kg dose volume. All standard agent preparations weremade fresh prior to their administration.

BxPC-3 cells were obtained and prepared as follows. The BxPC-3 pancreastumor cell line was received from American Type Culture Collection(ATCC, Manassas, Va.). Cultures were maintained in RPMI 1640 medium(Hyclone, Logan, Utah) supplemented with 5% fetal bovine serum. Thecells were housed in a 5% CO₂ atmosphere. The cultures were expanded intissue culture flasks at a 1:3 split ratio until a sufficient amount ofcells were harvested.

All experiments were conducted on female athymic nude mice (Hsd:AthymicNude-Foxn1nu) supplied by Harlan (Indianapolis, Ind.). Mice werereceived at four weeks of age, 12-15 grams in weight, and wereacclimated for seven days prior to handling. The mice were housed inmicroisolator cages (Lab Products, Seaford, Del.) and maintained underspecific pathogen-free conditions. All procedures were carried out underappropriate institutional guidelines for animal care.

BxPC-3 Human Pancreas Tumor Xenograft Model: Female athymic nude miceper treatment condition were inoculated subcutaneously in the rightflank with 0.1 ml of a 50% RPMI 1640/50% MatrigelTM (BD Biosciences,Bedford, Mass.) mixture containing a suspension of BxPC-3 tumor cells(approximately 5×10⁶ cells/mouse).

Seven days following inoculation, tumors were measured using calipersand tumor weight was calculated using the animal study managementsoftware, Study Director V.1.7.54k (Study Log). See Britten CD, et al.,“Enhanced antitumor activity of 6-hydroxymethylacylfulvene incombination with irinotecan and 5-fluorouracil in the HT29 human colontumor xenograft model”, Cancer Res 59:1049-1053, 1999. Eighty mice withtumor sizes of 93-172 mg were placed into eight groups of ten mice byrandom equilibration (Day 1). Body weights were recorded when the micewere randomized and were taken twice weekly thereafter in conjunctionwith tumor measurements, on each of Days 1, 4, 8, 11, 15, 18, 22, 26,30, 33, and 36.

ODSH, vehicle control (0.9% NaCl solution, referred to as saline),oxaliplatin, gemcitabine, and nab-paclitaxel were administered accordingto the dosing regimen described in Table 1. The study was terminatedwhen the vehicle control reached an endpoint of 1500 mg, on Day 36.Table 1, below, provides further details on the eight treatment groups.

TABLE 1 Treatment Agent group Treatment administered Dosing schedule andamount Route* 1 Vehicle control 0.9% saline Twice a day, Day 1 to Day 11IV Twice a day, Day 12 to Day 35 SC 2 ODSH 24 mg/kg Twice a day, Day 1to Day 11 IV Twice a day, Day 12 to Day 35 SC 3 Oxaliplatin 10 mg/kgOnce a week for 4 weeks (Days 1, 8, 15, 22) IV Gemcitabine 80 mg/kgEvery three days for 3 administrations (Days 26, 29, 32) IPNab-paclitaxel 15 mg/kg Every three days for 3 administrations (Days 26,29, 32) IV 4 Gemcitabine 80 mg/kg Every three days for 4 administrations(Days 1, 4, 7, 10) IP 5 ODSH/ 24 mg/kg Twice a day, Day 1 to Day 11 IVTwice a day, Day 12 to Day 35 SC Oxaliplatin 10 mg/kg Once a week for 4weeks (Days 1, 8, 15, 22) IV Gemcitabine 80 mg/kg Every three days for 4administrations (Days 26, 29, 32, 35) IP Nab-paclitaxel 15 mg/kg Everythree days for 3 administrations (Days 26, 29, 32) IV 6 ODSH 24 mg/kgTwice a day, Day 1 to Day 11 IV Twice a day, Day 12 to Day 35 SCGemcitabine 80 mg/kg Every three days for 4 administrations (Days 1, 4,7, 10) IP 7 Oxaliplatin 10 mg/kg Single administration (Day 1) IVGemcitabine 80 mg/kg Every three days for 3 administrations (Days 1, 4,7) IP 8 ODSH 24 mg/kg Twice a day, Day 1 to Day 8 IV Oxaliplatin 10mg/kg Single administration (Day 1) IV Gemcitabine 80 mg/kg Every threedays for 3 administrations (Days 1, 4, 7) IP *Agents were administeredby one of three routes: intravenous (IV), subcutaneous (SC), orintraperitoneal (IP).

Treatment for Groups 7 and 8 was ceased on Day 8 due to adverse effectsresulting from the treatment. The ODSH dosing route was modified fromintravenous to subcutaneous on Day 12, as a result of tail swelling andbruising. Gemcitabine and nab-paclitaxel were introduced into the dosingregimen of Groups 3 and 5 on Day 26.

Data and statistical analyses were performed as follows. Mean tumorgrowth inhibition (TGI) was calculated utilizing Formula A below (deathswere not included in the TGI calculations). TGI calculations wereperformed comparing tumor weights of Day 26 to Day 1, which capturesdata prior to the addition of gemcitabine and nab-paclitaxel to severalgroups, and Day 36 (final day of study) to Day 1.

$\begin{matrix}{{TGI} = {\left\lbrack {1 - \frac{\left( {{\overset{\_}{\chi}{Treated}_{({Final})}} = {\overset{\_}{\chi}{Treated}_{({{Day}\mspace{14mu} 1})}}} \right)}{\left( {{\overset{\_}{\chi}{Control}_{({Final})}} - {\overset{\_}{\chi}{Control}_{({{Day}\mspace{14mu} 1})}}} \right)}} \right\rbrack \times 100\%}} & {{Formula}\mspace{14mu} A}\end{matrix}$

All statistical analyses in the xenograft study were performed withGraphPad Prism® v4 software. Differences in Day 26 and 36 tumor weightswere confirmed using the Analysis of Variance (ANOVA) with the Tukey'sMultiple Comparison Test.

Results. The antitumor effects of ODSH administered as a single agent orin various combinations with one or more of oxaliplatin, gemcitabine,and nab-paclitaxel were evaluated.

The recorded tumor weights for experimental treatment groups 1 through 8are provided below in Tables 2 through 9. See also FIG. 1.

TABLE 2 PBS Control (0 mg/kg) Dose Route*: Intravenous/SubcutaneousFrequency: BID to end Day: Group 1 1 4 8 11 15 18 22 26 30 33 36 Mouse 1106 127 155 182 293 365 484 766 4161 4627 1,707 Mouse 2 114 140 195 190267 321 426 609 836 1,060 1379.03 Mouse 3 106 119 166 194 223 300 420635 1,067 1,306 1689.98 Mouse 4 93 103 120 160 237 338 449 681 1,1321,311 1565.56 Mouse 5 140 145 207 249 306 346 546 780 1,020 1,2301381.46 Mouse 6 114 118 138 154 218 273 344 557 722 986 1295.65 Mouse 7129 159 200 194 282 321 461 681 1,054 1,286 1,352 Mouse 8 130 122 134176 272 320 415 583 999 1,165 1,329 Mouse 9 142 153 164 184 318 333 465590 1,080 1,341 1,522 Mouse 10 172 194 259 285 377 505 665 915 1,3621,620 2,020 Mean 124.6 138.1 173.7 196.9 279.2 342.2 467.6 679.8 1,043.21,293.3 1,524.2 Median 121.6 133.7 164.8 186.9 276.8 327.3 454.9 657.91,060.4 1,296.2 1,451.9 Std Dev 22.78 26.24 41.72 40.36 48.10 62.4086.85 111.90 174.53 208.14 228.53 Std Err 7.20 8.30 13.19 12.76 15.2119.73 27.46 35.39 55.19 65.82 72.27 *ODSH dosed intravenously Days 1-11;dosed subcutaneously Days 12-end

TABLE 3 ODSH (24 mg/kg) Dose Route*: Intravenous/Subcutaneous Frequency:BID to end Day: Group 2 1 4 8 11 15 18 22 26 30 33 36 Mouse 1 143 144157 140 218 278 326 394 555 733 836 Mouse 2 129 133 148 203 264 352 515766 1,109 1,340 1,823 Mouse 3 140 162 218 149 206 278 358 509 697 9081,072 Mouse 4 114 98 118 288 340 489 622 894 1,083 1,218 1,450 Mouse 5116 110 144 142 224 291 360 478 756 999 1,153 Mouse 6 172 141 183 175263 288 400 529 893 961 1,150 Mouse 7 94 97 125 243 348 419 556 7701,201 1,309 1,654 Mouse 8 131 181 222 146 237 256 402 573 820 961 1,150Mouse 9 106 135 126 252 321 415 551 724 945 1,087 1,376 Mouse 10 125 130155 137 208 318 449 524 764 871 1,037 Mean 122.4 134.0 146.0 187.6 263.0338.5 453.8 616.2 882.3 1,038.6 1,270.1 Median 22.9 28.6 40.4 162.1250.0 304.7 425.4 551.3 856.4 979.8 1,151.5 Std Dev 7.24 9.03 12.7655.55 54.66 77.75 101.08 160.86 203.52 197.30 302.01 Std Err 17.57 17.2924.59 31.96 50.87 64.36 62.39 95.50 *ODSH dosed intravenously Days 1-11;dosed subcutaneously Days 12-end

TABLE 4 Oxaliplatin 10 mg/kg Dose Route: Intravenous Frequency: Wkly × 4(Day 1, 8, 15, 22) Gemcitabine 80 mg/kg Dose Route: IntraperitonealFrequency: Day 26, 39, 32 (Q3d × 3 starting Day 26) Nab/paclitaxel 15mg/kg Dose Route: Intravenous Frequency: Day 26, 29, 32 (2 × weeklystarting Day 26) Day: Group 3{circumflex over ( )} 1 4 8 11 15 18 22 2630 33 36 Mouse 1 113 144 154 202 271 300 392 570 725 FD FD Mouse 2 131135 219 184 250 269 322 397 393 MS MS Mouse 3 104 113 142 164 219 289379 487 595 FD FD Mouse 4 143 148 205 265 368 485 611 812 FD FD FD Mouse5 168 169 245 299 394 533 673 918 FD FD FD Mouse 6 128 114 167 201 282314 451 522 658 FD FD Mouse 7 96 126 161 211 263 362 432 629 687 FD FDMouse 8 116 136 205 243 308 461 666 824 926 FD FD Mouse 9 106 139 165222 301 371 522 600 706 MS MS Mouse 10 139 133 157 217 282 394 495 589689 MS MS Mean 124.5 135.7 182.0 220.6 293.7 377.6 494.4 635.0 672.2Median 121.9 135.7 165.8 213.6 282.0 366.4 473.1 594.5 688.0 Std Dev21.78 16.25 33.99 39.41 52.76 90.07 122.52 165.28 147.79 Std Err 6.895.14 10.75 12.46 16.68 28.48 38.74 52.26 52.25 {circumflex over( )}Beginning Day 26, groups 3 and 5 were taken off initial dosingregimen and the following dosing regimen was initiated: Gr 3:Gemcitabine (80 mg/kg IP, Q3d × 4 starting Day 26) + Nab-paclitaxel (15mg/kg IV, 2 × weekly starting Day 26) Gr 5: ODSH (24 mg/kg, IV BID) +Gemcitabine (80 mg/kg IP, Q3d × 4 starting Day 26) + Nab-paclitaxel (15mg/kg IV, 2 × weekly starting day 26)

TABLE 5 Gemcitabine 80 mg/kg Dose Route: Intraperitoneal Frequency: Q3d× 4 (Day 1, 4, 7, 10) Day: Group 4 1 4 8 11 15 18 22 26 30 33 36 Mouse 1146 155 180 234 300 389 500 647 992 1,158 1,267 Mouse 2 96 135 134 180274 304 404 583 826 1,008 1,265 Mouse 3 132 189 193 235 319 413 621 7971,121 1,367 1,573 Mouse 4 139 214 282 328 375 379 514 739 1,008 1,1321,368 Mouse 5 126 126 129 146 211 258 372 468 674 878 963 Mouse 6 104122 114 145 218 267 400 534 860 1,036 1,347 Mouse 7 107 131 130 119 181224 350 431 667 936 1,106 Mouse 8 111 133 125 150 228 269 332 389 589624 923 Mouse 9 116 146 161 202 245 441 592 730 1,150 1,275 1,596 Mouse10 166 176 175 242 288 376 509 628 856 1,030 1,482 Mean 124.3 152.5162.2 198.2 263.8 332.1 459.4 594.7 874.3 1,044.2 1,288.8 Median 121.1140.2 147.4 191.0 259.2 340.1 451.9 605.8 858.0 1,033.3 1,306.7 Std Dev21.73 30.57 50.04 63.22 58.46 76.00 101.69 138.55 193.10 209.66 234.90Std Err 6.87 9.67 15.83 19.99 18.49 24.03 32.16 43.81 61.06 66.30 74.28

TABLE 6 ODSH 24 mg/kg Dose Route*: Intravenous/Subcutaneous Frequency:BID to end Oxaliplatin 10 mg/kg Dose Route: Intravenous Frequency: Wkly× 4 (Day 1, 8, 15, 22) Gemcitabine 80 mg/kg Dose Route: IntraperitonealFrequency: Day 26, 29, 32, 35 (Q3d × 4 starting Day 26) Nab/paclitaxel15 mg/kg Dose Route: Intravenous Frequency: Day 26, 29, 32 (2 × weeklystarting Day 26) Day: Group 5{circumflex over ( )} 1 4 8 11 15 18 22 2630 33 36 Mouse 1 139 144 207 237 395 448 571 819 908 FD FD Mouse 2 103106 172 230 298 372 403 609 735 FD FD Mouse 3 160 162 166 205 279 337484 635 758 FD FD Mouse 4 97 113 108 113 160 173 225 313 359 FD FD Mouse5 125 134 174 163 245 299 421 576 772 FD FD Mouse 6 117 137 124 165 229259 334 416 552 MS MS Mouse 7 148 191 186 184 286 337 470 642 745 891828 Mouse 8 108 136 141 198 280 386 539 766 MS MS MS Mouse 9 133 177 214226 313 400 578 631 834 FD FD Mouse 10 111 119 156 173 219 229 333 457FD FD FD Mean 124.1 142.0 164.9 189.5 270.4 324.0 436.0 586.6 708.0891.1 827.5 Median 121.2 136.4 169.2 191.2 279.4 336.7 445.8 620.2 751.6891.1 877.5 Std Dev 20.52 27.53 34.05 37.97 62.97 84.34 115.23 154.52173.64 Std Err 6.49 8.71 10.77 12.01 19.91 26.67 36.44 48.86 61.39{circumflex over ( )}Beginning Day 26, groups 3 and 5 were taken offinitial dosing regimen and the following dosing regimen was initiated:Gr 3: Gemcitabine (80 mg/kg IP, Q3d × 4 starting Day 26) +Nab-paclitaxel (15 mg/kg IV, 2 × weekly starting Day 26) Gr 5: OD SH (24mg/kg, IV BID) + Gemcitabine (80 mg/kg IP, Q3d × 4 starting Day 26) +Nab-paclitaxel (15 mg/kg IV, 2 × weekly starting day 26) *ODSH dosedintravenously Days 1-11; dosed subcutaneously Days 12-end MS = MoribundSacrifice FD = Found Dead

TABLE 7 ODSH 24 mg/kg Dose Route*: Intravenous/Subcutaneous Frequency:BID to end Gemcitabine 80 mg/kg Dose Route: Intraperitoneal Frequency:Q3d × 4 Day 1, 4, 7, 10) Day: Group 6 1 4 8 11 15 18 22 26 30 33 36Mouse 1 160 169 194 135 203 279 407 562 780 893 1,115 Mouse 2 101 92 113151 188 194 309 374 497 578 862 Mouse 3 119 160 170 209 278 257 326 452571 822 997 Mouse 4 134 122 128 154 188 231 330 409 595 773 834 Mouse 5125 134 141 152 197 265 332 520 622 856 965 Mouse 6 97 102 117 156 203229 336 491 746 820 1,048 Mouse 7 137 121 166 171 205 256 389 510 704789 868 Mouse 8 151 174 203 255 331 484 635 886 1,069 1,289 1,365 Mouse9 111 120 123 130 149 199 273 342 490 631 833 Mouse 10 108 134 131 118239 191 323 543 757 843 1,051 Mean 124.2 132.7 148.7 163.0 218.1 258.5336.0 508.9 683.3 829.3 993.9 Median 121.7 128.1 136.1 153.3 203.0 243.2331.0 500.3 663.0 820.9 981.0 Std Dev 20.92 27.40 32.50 40.78 52.1285.24 101.65 151.31 171.22 189.79 164.57 Std Err 6.61 8.67 10.28 12.9016.48 26.96 32.15 47.85 54.15 60.02 52.04 *ODSH dosed intravenously Days1-11; dosed subcutaneously Days 12-end

TABLE 8 Oxaliplatin 10 mg/kg Dose Route: Intravenous Frequency: Day 1Gemcitabine 80 mg/kg Dose Route: Intraperitoneal Frequency: Q3d × 3 (Day1, 4, 7) Day: Group 7 1 4 8 11 15 18 22 26 30 33 36 Mouse 1 137 119 132110 MS MS MS MS MS MS MS Mouse 2 159 175 165 197 259 368 453 527 735 787877 Mouse 3 110 104 107 137 MS MS MS MS MS MS MS Mouse 4 110 114 110 117MS MS MS MS MS MS MS Mouse 5 135 117 81 FD FD FD FD FD FD FD FD Mouse 697 98 103 106 FD FD FD FD FD FD FD Mouse 7 152 173 222 217 314 389 541801 1,221 1,599 1,699 Mouse 8 121 117 113 FD FD FD FD FD FD FD FD Mouse9 119 146 150 173 272 299 424 510 715 871 1,107 Mouse 10 100 111 103 FDFD FD FD FD FD FD FD Mean 124.1 127.2 128.7 150.9 281.7 352.1 472.7612.4 890.1 1,085.5 1,227.7 Median 120.1 116.6 111.8 136.8 272.0 368.5452.8 526.9 734.8 870.9 1,107.0 Std Dev 21.16 27.63 41.13 44.59 29.0147.57 60.91 163.25 286.32 446.77 423.84 Std Err 6.69 8.74 13.01 16.8516.75 27.46 36.17 94.25 165.31 257.94 244.71 MS = Moribund Sacrifice FD= Found Dead

TABLE 9 ODSH 24 mg/kg Dose Route*: Intravenous/Subcutaneous Frequency:BID × 8 days Oxaliplatin 10 mg/kg Dose Route: Intravenous Frequency: Day1 Gemcitabine 80 mg/kg Dose Route: Intraperitoneal Frequency: Q3d × 3Day 1, 4, 7) Day: Group 8 1 4 8 11 15 18 22 26 30 33 36 Mouse 1 121 129127 FD FD FD FD FD FD FD FD Mouse 2 153 147 94 FD FD FD FD FD FD FD FDMouse 3 157 174 183 FD FD FD FD FD FD FD FD Mouse 4 110 102 81 FD FD FDFD FD FD FD FD Mouse 5 121 123 FD FD FD FD FD FD FD FD FD Mouse 6 110124 127 FD FD FD FD FD FD FD FD Mouse 7 99 82 121 FD FD FD FD FD FD FDFD Mouse 8 135 152 148 FD FD FD FD FD FD FD FD Mouse 9 136 159 215 196MS MS MS MS MS MS MS Mouse 10 99 118 122 FD FD FD FD FD FD FD FD Mean124.1 131.0 135.4 196.0 Median 121.0 126.3 127.0 196.0 Std Dev 20.7927.70 4.84 Std Err 6.57 8.76 13.95 FD = Found Dead MS = MoribundSacrifice *ODSH dosed intravenously Days 1-11; dosed subcutaneously Days12-end {circumflex over ( )}: Beginning Day 26, groups 3 and 5 weretaken off initial dosing regimen and the following dosing regimen wasinitiated: Gr 3: Gemcitabine (80 mg/kg IP, Q3d × 4 starting Day 26) +Nab-paclitaxel (15 mg/kg IV, 2 × weekly starting Day 26) Gr 5: OD SH (24mg/kg, IV BID) + Gemcitabine (80 mg/kg IP, Q3d × 4 starting Day 26) +Nab-paclitaxel (15 mg/kg IV, 2 × weekly starting day 26)

Tables 10 to 13 below show the body weights recorded for treatmentgroups 1 to 8 over the course of the experiment. See also FIG. 3.

TABLE 10 Day 4 Day 8 Day 1 % % Starting Average Average Weight AverageWeight Weight # Weight # Weight Loss or # Weight Loss or Group CompoundDosage Frequency Dose Route* (g) Mice (g) Mice (g) Gain Mice (g) Gain 1Vehicle  0 mg/kg BID to end Intravenous/ 20.86 10 20.86 10 20.48 −1.8210 20.67 −0.91 Control Subcutaneous 2 ODSH 24 mg/kg BID to endIntravenous/ 21.54 10 21.54 10 21.18 −1.67 10 21.18 −1.67 Subcutaneous3{circumflex over ( )} Oxaliplatin 10 mg/kg Wkly × 4 (Day Intravenous20.82 10 20.82 10 20.40 −2.02 10 20.65 −0.82 1, 8, 15, 22) 4 Gemcitabine80 mg/kg Q3d × 4 (Day Intraperitoneal 22.48 10 22.48 10 21.76 −3.20 1020.91 −6.98 1, 4, 7, 10) 5{circumflex over ( )} ODSH + 24 mg/kg + BID toend + Intravenous/ 21.11 10 21.11 10 20.40 −3.36 10 21.08 −0.14Oxaliplatin 10 mg/kg Wkly × 4 Subcutaneous (Day 1, 8, 15, 22) 6 ODSH +24 mg/kg + BID to end + Intravenous/ 20.95 10 20.95 10 20.31 −3.05 1018.92 −9.69 Gemcitabine 80 mg/kg Q3d × 4 Subcutaneous + (Day 1, 4, 7,Intravenous 10) 7 Oxaliplatin + 10 mg/kg + Day 1 + Intravenous + 20.5810 20.58 10 19.32 −6.12 10 17.17 −16.57 Gemcitabine 80 mg/kg Q3d × 3Intravenous (Day 1, 4, 7) 8 ODSH + 24 mg/kg + BID × 8 days +Intravenous + 20.96 10 20.96 10 18.88 −9.92 9 15.50 −26.05 Oxaliplatin +10 mg/kg + Day 1 + Intravenous + Gemcitabine 80 mg/kg Q3d × 3 (DayIntraperitoneal 1, 4, 7) {circumflex over ( )}Beginning Day 26, groups 3and 5 were taken off initial dosing regimen and the following dosingregimen was initiated: Gr 3: Gemcitabine (80 mg/kg IP, Q3d × 4 startingDay 26) + Nab-paclitaxel (15 mg/kg IV, 2 × weekly starting Day 26) Gr 5:ODSH (24 mg/kg, IV BID) + Gemcitabine (80 mg/kg IP, Q3d × 4 starting Day26) + Nab-paclitaxel (15 mg/kg IV, 2 × weekly starting day 26) *ODSHdosed intravenously Days 1-11; dosed subcutaneously Days 12-end

TABLE 11 Day 11 Day 15 Day 18 % % % Average Weight Average WeightAverage Weight # Weight Loss or # Weight Loss or # Weight Loss or GroupCompound Dosage Frequency* Dose Route Mice (g) Gain Mice (g) Gain Mice(g) Gain 1 Vehicle  0 mg/kg BID to end Intravenous/ 10 20.45 −1.97 1020.10 −3.64 10 20.36 −2.40 Control Subcutaneous 2 ODSH 24 mg/kg BID toend Intravenous/ 10 20.95 −2.74 10 21.08 −2.14 10 21.04 −2.32Subcutaneous 3{circumflex over ( )} Oxaliplatin 10 mg/kg Wkly × 4 (DayIntravenous 10 20.12 −3.36 10 20.81 −0.05 10 20.45 −1.78 1, 8, 15, 22) 4Gemcitabine 80 mg/kg Q3d × 4 (Day Intraperitoneal 10 20.52 −8.72 1021.70 −3.47 10 23.15 2.98 1, 4, 7, 10) 5{circumflex over ( )} ODSH + 24mg/kg + BID to end + Intravenous/ 10 19.79 −6.25 10 19.87 −5.87 10 19.49−7.67 Oxaliplatin 10 mg/kg Wkly × 4 Subcutaneous + (Day 1, 8, 15,Intravenous 22) 6 ODSH + 24 mg/kg + BID to end + Intravenous/ 10 18.14−13.41 10 18.84 −10.07 10 20.68 −1.29 Gemcitabine 80 mg/kg Q3d × 4Subcutaneous + (Day 1, 4, 7, 10) Intravenous 7 Oxaliplatin + 10 mg/kg +Day 1 + Q3d × 3 Intravenous + 7 17.17 −16.56 3 22.03 7.06 3 23.37 13.54Gemcitabine 80 mg/kg (Day 1, 4, 7) Intravenous 8 ODSH + 24 mg/kg + BID ×8 days + Intravenous + 1 14.80 −29.39 0 0 Oxaliplatin + 10 mg/kg + Day1 + Q3d × 3 Intravenous + Gemcitabine 80 mg/kg (Day 1, 4, 7)Intraperitoneal {circumflex over ( )}Beginning Day 26, groups 3 and 5were taken off initial dosing regimen and the following dosing regimenwas initiated: Gr 3: Gemcitabine (80 mg/kg IP, Q3d × 4 starting Day26) + Nab-paclitaxel (15 mg/kg IV, 2 × weekly starting Day 26) Gr 5:ODSH (24 mg/kg, IV BID) + Gemcitabine (80 mg/kg IP, Q3d × 4 starting Day26) + Nab-paclitaxel (15 mg/kg IV, 2 × weekly starting day 26) *ODSHdosed intravenously Days 1-11; dosed subcutaneously Days 12-end

TABLE 12 Day 22 Day 26 Day 30 % % % Average Weight Average WeightAverage Weight # Weight Loss or # Weight Loss or # Weight Loss or GroupCompound Dosage Frequency* Dose Route Mice (g) Gain Mice (g) Gain Mice(g) Gain 1 Vehicle  0 mg/kg BID to end Intravenous/ 10 20.81 −0.24 1020.93 0.34 10 21.28 2.01 Control Subcutaneous 2 ODSH 24 mg/kg BID to endIntravenous/ 10 21.56 0.09 10 22.14 2.79 10 22.60 4.92 Subcutaneous3{circumflex over ( )} Oxaliplatin 10 mg/kg Wkly × 4 (Day Intravenous 1021.27 2.16 10 20.96 0.67 8 18.08 −13.18 Gemcitabine 80 mg/kg 1, 8, 15,22) Intraperitoneal Nab-paclitaxel 15 mg/kg Day 26, 29, 32 IntravenousDay 26, 29, 32 4 Gemcitabine 80 mg/kg Q3d × 4 (Day Intraperitoneal 1024.22 7.74 10 24.27 7.96 10 24.19 7.61 1, 4, 7, 10) 5{circumflex over( )} ODSH + 24 mg/kg + BID to end + Intravenous/ 10 20.47 −3.03 10 20.51−2.84 8 18.16 −13.96 Oxaliplatin 10 mg/kg Wkly × 4 Subcutaneous +Gemcitabine 80 mg/kg (Day 1, 8, 15, 22) Intravenous Nab-paclitaxel 15mg/kg Day 26, 29, 32, 35 Intraperitoneal Day 26, 29, 32 Intravenous 6ODSH + 24 mg/kg + BID to end + Intravenous/ 10 21.64 3.29 10 22.79 8.7810 22.36 6.73 Gemcitabine 80 mg/kg Q3d × 4 (Day 1, Subcutaneous + 4, 7,10) Intravenous 7 Oxaliplatin + 10 mg/kg + Day 1 + Q3d × 3 Intravenous +3 23.67 15.00 3 23.73 15.32 3 23.90 16.13 Gemcitabine 80 mg/kg (Day 1,4, 7) Intravenous 8 ODSH + 24 mg/kg + BID × 8 days + Intravenous + 0 0 00 0 Oxaliplatin + 10 mg/kg + Day 1 + Intravenous + Gemcitabine 80 mg/kgQ3d × 3 (Day 1, Intraperitoneal 4, 7) {circumflex over ( )}Beginning Day26, groups 3 and 5 were taken off initial dosing regimen and thefollowing dosing regimen was initiated: Gr 3: Gemcitabine (80 mg/kg IP,Q3d × 4 starting Day 26) + Nab-paclitaxel (15 mg/kg IV, 2 × weeklystarting Day 26) Gr 5: OD SH (24 mg/kg, IV BID) + Gemcitabine (80 mg/kgIP, Q3d × 4 starting Day 26) + Nab-paclitaxel (15 mg/kg IV, 2 × weeklystarting day 26) *ODSH dosed intravenously Days 1-11; dosedsubcutaneously Days 12-end

TABLE 13 Day 22 Day 26 % % Average Weight Average Weight Weight Loss orWeight Loss or Group Compound Dosage Frequency Dose Route* # Mice (g)Gain # Mice (g) Gain 1 Vehicle  0 mg/kg BID to end Intravenous/ 10 21.493.02 10 21.46 2.88 Control Subcutaneous 2 ODSH 24 mg/kg BID to endIntravenous/ 10 22.53 4.60 10 22.46 4.27 Subcutaneous 3{circumflex over( )} Oxaliplatin 10 mg/kg Wkly × 4 (Day 1, 8, 15, 22) Intravenous 0 0 00 Gemcitabine 80 mg/kg Day 26, 29, 32 Intraperitoneal Nab-paclitaxel 15mg/kg Day 26, 29, 32 Intravenous 4 Gemcitabine 80 mg/kg Q3d × 4 (Day 1,4, 7, 10) Intraperitoneal 10 24.41 8.59 10 24.21 7.70 5{circumflex over( )} ODSH + 24 mg/kg + BID to end + Wkly × 4 Intravenous/ 1 19.10 −9.521 17.20 −18.52 Oxaliplatin 10 mg/kg (Day 1, 8, 15, 22) Subcutaneous +Gemcitabine 80 mg/kg Day 26, 29, 32, 35 Intravenous Nab-paclitaxel 15mg/kg Day 26, 29, 32 Intraperitoneal Intravenous 6 ODSH + 24 mg/kg + BIDto end + Q3d × 4 Intravenous/ 10 22.29 6.40 10 22.03 5.16 Gemcitabine 80mg/kg (Day 1, 4, 7, 10) Subcutaneous + Intravenous 7 Oxaliplatin + 10mg/kg + Day 1 + Q3d × 3 Intravenous + 3 23.87 15.97 3 24.37 18.40Gemcitabine 80 mg/kg (Day 1, 4, 7) Intravenous 8 ODSH + 24 mg/kg + BID ×8 days + Intravenous + 0 0 Oxaliplatin + 10 mg/kg + Day 1 +Intravenous + Gemcitabine 80 mg/kg Q3d × 3 (Day 1, 4, 7) Intraperitoneal{circumflex over ( )}Beginning Day 26, groups 3 and 5 were taken offinitial dosing regimen and the following dosing regimen was initiated:Gr 3: Gemcitabine (80 mg/kg IP, Q3d × 4 starting Day 26) +Nab-paclitaxel (15 mg/kg IV, 2 × weekly starting Day 26) Gr 5: ODSH (24mg/kg, IV BID) + Gemcitabine (80 mg/kg IP, Q3d × 4 starting Day 26) +Nab-paclitaxel (15 mg/kg IV, 2 × weekly starting day 26) *ODSH dosedintravenously Days 1-11; dosed subcutaneously Days 12-end

Efficacy was assessed by comparison of tumor weights at Day 26 and 36against Day 1. Day 26 was chosen to assess data prior to the addition ofgemcitabine and nab-paclitaxel to groups 3 and 5. Day 36 was assessed asthe last day of the study.

Tables 14 and 15, below, show the tumor weight and percent tumor growthinhibition (%TGI) for all treatment groups relative to Group 1 (thevehicle control group) at Day 26 and Day 36. See also FIG. 1 and FIG. 2.

TABLE 14 Day 26 Tumor Weight Day 26 Group Treatment¹ N Dose Schedule(mg) % TGI (n) Deaths² 1 Vehicle 10 — BID to end 679.8 ± 35.4 — 0 2 ODSH10 24 mg/kg BID to end 616.2 ± 50.9 11.5 0 (10/10) 3 Oxaliplatin 10 10mg/kg Wkly × 4 635.0 ± 52.3 8.0 0 (10/10) 4 Gemcitabine 10 80 mg/kg Q3D× 4 594.7 ± 43.8 15.3 0 (10/10) 5 ODSH 10 24 mg/kg BID to end 586.6 ±48.9 16.7 0 Oxaliplatin 10 mg/kg Wkly × 4 (10/10) 6 ODSH 10 24 mg/kg BIDto end 508.9 ± 47.9 30.7 0 Gemcitabine 80 mg/kg Q3D × 4 (10/10) 7Oxaliplatin 10 10 mg/kg Day 1 612.4 ± 94.3 15.5 7 Gemcitabine 80 mg/kgQ3D × 3  (3/10) 8 ODSH 10 24 mg/kg BID to end — — 10 Oxaliplatin 10mg/kg Day 1 Gemcitabine 80 mg/kg Q3D × 3 ¹Treatment administered untilDay 26 ²Total deaths on or before Day 26

TABLE 15 Day 36 Tumor Day 36 Weight % TGI Group Treatment¹ n DoseSchedule (mg) (n) Deaths² 1 Vehicle 10 — BID to end 1524.2 ± 72.3 — 0 2ODSH 10 24 mg/kg BID to end 1270.1 ± 95.5 18.2 0 (10/10) 3 Oxaliplatin10 10 mg/kg 2 × wkly × 4 — — 10 Gemcitabine 80 mg/kg Day nab-paclitaxel15 mg/kg 26, 29, 32 Day 26, 29, 32 4 Gemcitabine 10 80 mg/kg Q3D × 41288.8 ± 74.3 16.8 0 (10/10)

TABLE 15 Day 36 Tumor Day 36 Weight % TGI Group Treatment¹ n DoseSchedule (mg) (n) Deaths² 5 ODSH 10 24 mg/kg BID to end 827.5 51.4 9Oxaliplatin 10 mg/kg 1 × Wkly × (n = 1) (1/10) Gemcitabine 80 mg/kg 4Day nab-paclitaxel 15 mg/kg 26, 29, 32, 35 Day 26, 29, 32 6 ODSH 10 24mg/kg BID to end 993.9 ± 52.0 37.9 0 Gemcitabine 80 mg/kg Q3D × 4(10/10)  7 Oxaliplatin 10 10 mg/kg Day 1 1227.7 ± 244.7 22.5 7Gemcitabine 80 mg/kg Day 1, 4, 7 (3/10) 8 ODSH 10 24 mg/kg BID to end —— 10 Oxaliplatin 10 mg/kg Day 1 Gemcitabine 80 mg/kg Day 1, 4, 7¹Treatment administered until Day 35 ²Total deaths on or before Day 36

Mean tumor weight in the vehicle control group (Group 1) reached 679.8mg by Day 26 and 1524.2 mg by Day 36. Six of ten tumors demonstratedsome level of necrosis; however, this is attributed to the normalprogression of this tumor xenograft model. Tumor necrosis was firstobserved on Day 30. A maximum weight loss of 3.6% was observed at Day15. The mice recovered their weight by Day 26. Two of ten micedemonstrated slightly bruised tails, first observed on Day 11.

Mean tumor weight in the group receiving ODSH at 24 mg/kg (Group 2)reached 616.2 mg by Day 26 and 1270.1 mg by Day 36. This treatmentresulted in a TGI of 11.5% on Day 26 and 18.2% on Day 36, relative toDay 1. No significant difference in tumor weight was observed on Day 26when compared to vehicle control, however by Day 36 a significantdifference in tumor weight was seen, in Group 2 relative to the vehiclecontrol group. Three of ten tumors demonstrated some level of necrosis;however, this is attributed to the normal progression of this tumorxenograft model. Tumor necrosis was first observed on Day 30. A maximumweight loss of 2.7% was reached on Day 11. The mice recovered theirweight by Day 22. All ten mice in this group demonstrated bruising onthe tails or abdomen, at the site of injection. This was first observedon Day 8 for the tails and Day 15 for the abdomens. One of the ten micealso demonstrated swelling of the tail, first observed on Day 11.

Oxaliplatin 10 mg/kg or gemcitabine 80 mg/kg, and nab-paclitaxel 15mg/kg (Group 3): The initial regimen of oxaliplatin alone reached a meantumor weight of 635.0 mg by Day 26, prior to the addition of gemcitabineand nab-paclitaxel to the dosing regimen. This group produced a TGI of8.0% on Day 26, when compared to vehicle control. No significantdifference in tumor weight on Day 26 was observed when compared tovehicle control. One mouse exhibited a bruised tail, first observed onDay 11. Three of ten tumors demonstrated some level of necrosis;however, this is attributed to the normal progression of this tumorxenograft model. Tumor necrosis was first observed on Day 26.

Following data collection on Day 26, the combination treatment regimenof gemcitabine and nab-paclitaxel was initiated. This regimen proved tobe toxic following the initial oxaliplatin alone treatment. No efficacydata could be reported for the triple combination.

Gemcitabine 80 mg/kg (Group 4) reached a mean tumor weight of 594.7 mgby Day 26 and 1288.8 mg by Day 36. This treatment resulted in a TGI of15.3% on Day 26 and 16.8% on Day 36, when compared to vehicle control.No significant difference in tumor weight was observed on Day 26 or Day36 when compared to vehicle control. Four of the ten tumors demonstratedsome level of necrosis; however, this is attributed to the normalprogression of this tumor xenograft model. Tumor necrosis was firstobserved on Day 26. A maximum weight loss of 8.7% was reached on Day 11.The mice recovered their weight by Day 18.

ODSH 24 mg/kg and oxaliplatin 10 mg/kg or ODSH 24mg/kg, gemcitabine 80mg/kg, and nab-paclitaxel 15 mg/kg (Group 5): The initial treatmentcombination of ODSH and oxaliplatin reached a mean tumor weight of 586.6mg by Day 26. This treatment resulted in a TGI of 16.7% on Day 26 whencompared to vehicle control. No significant difference in tumor weightwas observed on Day 26 when compared to vehicle control, ODSH (Group 2),or oxaliplatin (Group 3). All ten mice in this group demonstratedincreased bruising on the tails or abdomen, at the site of injection.This was first observed on Day 4 for the tails and Day 15 for theabdomens. Two of the ten mice also demonstrated swelling of the tail,first observed on Day 4. Three of the ten mice demonstrated somediscoloration of the skin, first observed on Day 11.

The triple combination of ODSH, gemcitabine, and nab-paclitaxel,initiated on Day 26, resulted in increased toxicity following theinitial treatment regimen of ODSH and oxaliplatin. No statisticalanalysis could be performed on Day 36 because only one mouse remained inthis group to Day 36 with a tumor size of 827.5 mg (TGI=51.4%).

ODSH 24 mg/kg and gemcitabine 80 mg/kg (Group 6) reached a mean tumorweight of 508.9 mg by Day 26 and 993.9 mg by Day 36. This treatmentresulted in a TGI of 30.7% on Day 26 and 37.9% on Day 36 when comparedto vehicle control. No significant difference in tumor weight wasobserved on Day 26 when compared to vehicle control, ODSH (Group 2), orgemcitabine (Group 4). A significant decrease in tumor weight was seenon Day 36 (P<0.05) when compared to vehicle control; however, nosignificant difference in tumor weights resulted when compared to ODSH(Group 2) or gemcitabine (Group 4). One of ten tumors demonstrated somelevel of necrosis; however, this is attributed to the normal progressionof this tumor xenograft model. Tumor necrosis was first observed on Day30. A maximum weight loss of 13.4% was reached on Day 11. The micerecovered their weight by Day 22. All ten mice in this groupdemonstrated bruising on the tails or abdomen, at the site of injection.This was first observed on Day 4 for the tails and Day 15 for theabdomens. Two of the ten mice also demonstrated swelling of the tail,first observed on Day 4. One of ten mice demonstrated discoloration ofthe skin, first observed on Day 9. Two of ten mice demonstrated dryskin, first observed on Day 9.

Oxaliplatin 10 mg/kg and gemcitabine 80 mg/kg (Group 7) reached a meantumor weight of 612.4 mg by Day 26 and 1227.7 mg by Day 36. This groupproduced a TGI of 15.5% on Day 26 (n=3) and 22.5% on Day 36 (n=3), whencompared to the vehicle control. No significant difference in tumorweight was observed on Day 26, when compared to vehicle control,oxaliplatin (Group 3), or gemcitabine (Group 4). No significantdifference in tumor weight was observed on Day 36 when compared tovehicle control or gemcitabine (Group 4). One of ten tumors demonstratedsome level of necrosis; this is attributed to the natural progression ofthe xenograft model. Tumor necrosis was first observed on Day 30. Amaximum weight loss of 16.6% was reached on Day 8. The mice recoveredtheir weight by Day 15 following cessation of gemcitabine treatment.This treatment regimen proved to be toxic. Mice were found dead on Days10, 11, and 14, and moribund sacrificed on Days 11 and 12. Two of tenmice in this group demonstrated bruising on the tails. This was firstobserved on Day 4.

ODSH 24 mg/kg, oxaliplatin 10 mg/kg, and gemcitabine 80 mg/kg (Group 8)could not be assessed for efficacy due to the toxicity of the regimendriven by the oxaliplatin and gemcitabine doses.

Treatment with ODSH alone was well-tolerated although some bruising andswelling at the site of injections occurred. Therefore, the dosing routewas changed to subcutaneous injection at Day 12. The combinationtreatments of ODSH and gemcitabine and ODSH and oxaliplatin weretolerated. Conversely, treatment combination regimens that includedgemcitabine with oxaliplatin or gemcitabine and nab-paclitaxel resultedin toxicity.

The combination of ODSH and gemcitabine resulted in the best efficacy atDay 26 and Day 36. On both comparison days, the combination of ODSH andgemcitabine resulted in notably lower tumor weights than gemcitabinealone. The tumor weights of mice treated with ODSH and gemcitabine werestatistically significantly lower than tumor weights in the control(saline alone) group on Day 36. See FIG. 2.

The addition, on Day 26 of the study, of gemcitabine and nab-paclitaxelto the oxaliplatin regimen in Groups 3 and 5 demonstrated severetoxicity that led to the death of many animals. It is unclear whetherthese toxicities were due to the combined treatment with oxaliplatin,gemcitabine and nab-paclitaxel, or residual toxicity related to theadministration of oxaliplatin.

6.2. Example 2 ODSH, a PF4-Interacting Heparinoid, Enhances the Efficacyof Carboplatin in an In Vivo Murine Xenograft Model of Human OvarianCancer

This example demonstrates that adjunctive administration of ODSHenhances the efficacy of carboplatin against human ovarian tumorsgrowing as xenografts in athymic nude mice.

Materials & methods. ODSH (50 mg/mL stock concentration) was made byPyramid Laboratories, Inc. and stored at room temperature until use. Asdescribed further below, ODSH was administered either intravenously(IV), at a dose of 48 mg/kg and volume of 10 mL/kg, or subcutaneously(SC) at a dose of 24 mg/kg and volume of 5 mL/kg. Carboplatin obtainedfrom a clinical supplier was stored at 4° C. until use. Carboplatin wasadministered by intraperitoneal injection (IP) at a dose of 80 mg/kg andvolume of 10 mL/kg. Saline solution (0.9% NaCl) was used as a vehiclecontrol for ODSH and administered by the same routes and in the samevolumes as ODSH.

Human ovarian cancer cell line A2780 was used in the murine xenograftexperiments as follows. Approximately 5×10⁶ A2780 cells were used permouse, injected subcutaneously into the right flank in 0.1 mL of 50%Matrigel/50% media. The study was initiated when tumors reached a sizeof 90-130 mg. 40 mice were used, 10 per treatment regimen. The mice werenormal athymic female mice, aged 6-7 weeks, housed in microisolatorcages and maintained under pathogen-free conditions. Tumor and bodyweights were measured three times per week.

ODSH, oxaliplatin, and the vehicle control (0.9% NaCl) were administeredaccording to the dosing schedule described in the table below. Fourtreatment groups were studied.

TABLE A Treatment regimens Agent Treat- admin- ment istered, groupTreatment and amount Dosing schedule Route* 1 Vehicle 0.9% saline Days1, 8, 15 IV control Twice a day, until end of SC study on days withoutIV dosing 2 ODSH 48 mg/kg Days 1, 8, 15 IV 24 mg/kg Twice a day untilend of SC study on days without IV dosing 3 Carboplatin 80 mg/kg Days 1,8, 15 IP 4 ODSH 48 mg/kg Days 1, 8, 15 admin- IV istered immediatelyafter carboplatin 24 mg/kg Twice a day until end of SC study on dayswithout IV dosing Carboplatin 80 mg/kg Days 1, 8, 15 IP *IV =intravenous, SC = subcutaneous, IP = intraperitoneal.

Results. As shown in FIG. 4, ODSH does not reduce or counteract theanti-neoplastic effect of carboplatin. On the contrary, adjunctiveadministration of ODSH with carboplatin (FIG. 4, triangles) caused asignificant reduction in the weights of tumors of human ovarian cancercells, as compared to the reduction in tumor weights seen in micereceiving only carboplatin (FIG. 4, squares). Furthermore, mice treatedwith ODSH adjunctive to carboplatin had an increase in body weight ascompared to mice receiving only carboplatin. See FIG. 6, trianglesversus squares. Thus, ODSH not only increased the efficacy ofcarboplatin, it also contributed to improved body weights, indicative ofa positive effect on constitutional side effects (e.g., loss ofappetite, weight loss) often seen with chemotherapeutic treatmentregimens.

Recorded tumor weights (in mg) for mice in experimental treatment groups1 through 4 are provided in the tables below. Tumor weights weremeasured on days 1, 4, 6, 8, 11, 14, 18, 21, 25 and 28 as indicated.

Group 1 Vehicle Dose Intravenous + Subcutaneous Control Route: QD Day 1,8, 15, 22 (IV) + BID Days 2-7, 9-14, 16-20, 23-27 (SC) 0 mg/kgFrequency: Day: 1 4 6 8 11 14 18 21 25 28 Mouse 1 90 157 270 438 637 8161,166 1,513 2,045 2,566 Mouse 2 97 161 250 368 627 1,103 1,728 2,722 TSTS Mouse 3 101 131 164 340 435 790 1,199 1,437 2,251 3,542 Mouse 4 104184 295 435 591 975 1,486 2,231 TS TS Mouse 5 104 148 285 403 710 1,0531,797 2,313 TS TS Mouse 6 108 145 259 465 764 1,157 1,487 2,053 2,5473,425 Mouse 7 108 202 397 629 1047 1,330 2,138 3,480 TS TS Mouse 8 11267 303 391 725 1,080 1,563 2,051 2,940 4,020 Mouse 9 122 181 350 508 7411,171 1,825 2,600 TS TS Mouse 10 126 171 277 396 663 1,009 1,595 1,9912,388 3,246 Mean 107.3 164.6 285.0 437.3 694.0 1,048.5 1,598.3 2,239.12,434.1 3,359.7 Median 106.0 164.0 281.0 419.0 686.7 1,066.3 1,579.12,142.0 2,387.7 3,424.7 Std Dev 10.67 21.07 61.44 82.88 156.26 162.46292.64 597.23 337.54 528.10 Std Err 3.37 6.66 19.43 26.21 49.41 51.3792.54 188.86 150.95 236.17

Group 2 PGX ODSH Dose Route: Intravenous 48 mg/kg Frequency: QD Day 1,8, 15, 22 PGX ODSH Dose Route: Subcutaneous 24 mg/kg Frequency: BID Days2-7, 9-14, 16-20, 23-27 Day: 1 4 6 8 11 14 18 21 25 28 Mouse 1 90 121241 373 635 1,084 1,615 2,642 TS TS Mouse 2 96 117 171 315 491 815 1,1571,681 2,423 3,129 Mouse 3 101 138 201 296 529 762 1,422 2,025 TS TSMouse 4 104 159 309 449 816 1,303 1,796 2,692 TS TS Mouse 5 104 111 215321 366 670 1,150 1,467 2,414 3,909 Mouse 6 107 195 347 639 910 1,4991,836 2,639 TS TS Mouse 7 109 131 264 413 653 1,032 1,568 1,852 2,4643,784 Mouse 8 112 173 235 411 536 868 1,714 1,880 2,599 3,252 Mouse 9120 124 139 183 349 559 891 1,289 1,817 2,671 Mouse 10 126 200 282 422811 1,253 1,869 2,560 TS TS Mean 107.0 146.8 240.3 382.2 609.6 984.41,501.7 2,072.6 2,343.5 3,349.0 Median 105.5 134.2 238.2 392.0 585.4949.7 1,591.5 1952.5 2,422.8 3,251.6 Std Dev 10.61 32.80 62.97 119.75191.32 302.40 336.20 525.76 303.65 505.10 Std Err 3.36 10.37 19.91 37.8760.50 95.63 106.32 166.26 135.80 225.89 FD = Found Dead TS = TerminalSacrifice

Group 3 Carboplatin Dose Route: Intraperitoneal 80 mg/kg Frequency: QDDay 1, 8, 15 Day: 1 4 6 8 11 14 18 21 25 28 Mouse 1 94 137 200 419 517587 942 1,218 1,921 2,461 Mouse 2 96 108 157 245 442 706 730 941 TS TSMouse 3 102 109 169 262 402 640 671 1,040 1,246 1,739 Mouse 4 104 108114 148 103 96 84 112 TS TS Mouse 5 105 175 264 466 539 875 1,349 1,7902,403 2,877 Mouse 6 106 199 239 358 499 799 1,218 1,596 2,127 2,599Mouse 7 110 142 223 371 419 628 713 834 1,109 1,539 Mouse 8 113 163 300605 813 1,093 1,806 2,675 TS TS Mouse 9 116 147 208 306 555 795 1,1601,626 TS TS Mouse 10 127 194 256 386 542 975 1,168 1,439 TS TS Mean107.3 148.2 213.0 356.5 483.1 719.3 984.1 1,327.3 1,761.3 2,243.0 Median105.6 144.5 215.6 364.2 508.5 750.3 1,051.0 1,328.6 1,921.0 2,461.0 StdDev 9.84 34.21 55.78 127.36 176.21 271.33 468.66 680.53 561.55 575.98Std Err 3.11 10.82 17.64 40.27 55.72 85.80 148.20 215.20 251.13 257.59

Group 4 PGX ODSH Dose Route: Intravenous 48 mg/kg Frequency: QD Day 1,8, 15, 22 PGX ODSH Dose Route: Subcutaneous 24 mg/kg Frequency: BID Days2-7, 9-14, 16-20, 23-27 Carboplatin Dose Route: Intraperitoneal 80 mg/kgFrequency: QD Day 1, 8, 15 Day: 1 4 6 8 11 14 18 21 25 28 Mouse 1 94 108155 210 240 319 444 555 740 1,053 Mouse 2 95 140 173 231 342 414 614 7241,307 1,795 Mouse 3 103 107 132 141 146 153 246 406 TS TS Mouse 4 103153 179 FD FD FD FD FD FD FD Mouse 5 105 114 130 129 110 86 90 88 TS TSMouse 6 105 149 180 233 413 528 1,036 1,222 TS TS Mouse 7 110 130 140170 192 213 264 463 TS TS Mouse 8 113 168 238 394 612 1,027 1,732 2,033TS TS Mouse 9 114 134 149 186 229 458 725 887 1,198 1,501 Mouse 10 127139 147 166 271 388 429 471 629 910 Mean 107.2 134.2 162.9 206.7 284.0398.3 619.9 761.1 968.6 1,314.9 Median 105.2 136.4 151.9 185.7 240.4388.3 444.3 554.7 968.8 1,277.2 Std Dev 9.73 19.97 32.39 79.09 154.07276.98 504.37 574.83 334.13 407.49 Std Err 3.08 6.32 10.24 26.36 51.3692.33 168.12 191.61 167.06 203.75 FD = Found Dead TS = TerminalSacrifice

Recorded individual body weights (in grams) for mice in experimentaltreatment groups 1 through 4 are provided in the tables below. Bodyweights were measured on days 1, 4, 6, 8, 11, 14, 18, 21, 25 and 28 asindicated.

Individual weights (g) 1st 2nd Weight 3rd Weight 4th Weight Weight Day 4Day 6 Day 8 Day 1 Weight Weight Weight Weight Weight change Weightchange Weight change Groups Compound Dosage Route Schedule Mouse (g) (g)(%) (g) (%) (g) (%) Group 1 Vehicle  0 mg/kg Intravenous QD Days 1, 8, 120.5 21.3 3.9 20.9 2.0 21.5 4.9 Control Subcutaneous 15, 22 2 19.8 20.74.9 20.0 1.0 20.4 3.0 BID Days 2-7, 9-14, 3 23.1 24.5 6.1 24.3 5.2 23.83.0 16-20, 23-27 4 21.3 21.5 0.9 21.2 −0.5 22.2 4.2 5 19.0 19.1 0.5 19.10.5 19.3 1.6 6 22.1 22.7 2.7 23.1 4.5 23.6 6.8 7 22.1 25.5 15.4 25.013.1 24.3 10.0 8 21.6 21.7 0.5 21.1 −2.3 21.4 −0.9 9 22.1 22.2 0.5 21.6−2.3 21.7 −1.8 10 22.7 22.9 0.9 23.2 2.2 23.4 3.1 Group 2 PGX ODSH 48mg/kg Intravenous QD Days 2, 8, 1 22.1 22.9 3.6 23.2 5.0 23.2 5.0 PGXODSH 24 mg/kg Subcutaneous 15, 22 2 20.3 20.6 1.5 20.6 1.5 21.4 5.4 BIDDays 2-7, 9-14, 3 21.0 20.9 −0.5 21.2 1.0 21.6 2.9 16, 20, 23-27 4 21.821.4 −1.8 21.5 −1.4 22.3 2.3 5 21.6 23.0 6.5 24.5 13.4 25.1 16.2 6 21.421.8 1.9 22.0 2.8 22.6 5.6 7 19.8 19.6 −1.0 20.2 2.0 20.9 5.6 8 19.820.9 5.6 21.3 7.6 22.2 12.1 9 20.9 23.0 10.0 22.6 8.1 22.3 6.7 10 19.719.2 −2.5 19.2 −2.5 20.1 2.0 Group 3 Carboplatin 80 mg/kgIntraperitoneal QD 1, 8, 15 1 20.3 19.1 −5.9 19.4 −4.4 20.1 −1.0 2 21.621.1 −2.3 20.8 −3.7 21.3 −1.4 3 22.2 21.8 −1.8 21.5 −3.2 22.3 0.5 4 23.222.6 −2.6 22.5 −3.0 23.7 2.2 5 20.6 19.5 −5.3 19.6 −4.9 20.7 0.5 6 22.621.6 −4.4 22.9 1.3 24.0 6.2 7 21.7 21.0 −3.2 21.4 −1.4 22.0 1.4 8 20.721.1 1.9 21.5 3.9 23.8 5.3 9 22.4 22.3 −0.4 22.3 −0.4 22.9 2.2 10 19.718.6 −5.6 19.1 −3.0 20.0 1.5 Group 4 PGX OGSH 48 mg/kg Intravenous QDDays 1, 8, 1 21.0 21.4 1.9 22.0 4.8 22.8 8.6 PGX OGSH 24 mg/kgSubcutaneous 15, 22 2 22.4 22.1 −1.3 22.8 1.8 23.5 4.9 Carboplatin 80mg/kg Intraperiotoneal BID Days 2-7, 9-14, 3 21.6 21.3 −1.4 20.8 −3.722.0 1.9 16-20, 23-27 4 21.2 18.0 −15.1 15.2 −28.3 FD FD QD Days 1, 8,15 5 21.9 21.5 −1.8 22.4 2.3 23.2 5.9 6 20.5 20.0 −2.4 21.1 2.9 22.0 7.37 20.3 19.7 −3.0 20.1 −1.0 20.9 3.0 8 24.0 24.3 1.3 24.1 0.4 24.9 3.7 921.1 20.6 −2.4 21.0 −0.5 22.2 5.2 10 25.0 24.8 −0.8 25.7 2.8 25.6 2.4 FD= Found Dead TS = Terminal Sacrifice

Individual weights (g) 5th Weight 6th Weight Day 11 Day 14 Weight WeightWeight change Weight change Groups Compound Dosage Route Schedule Mouse(g) (%) (g) (%) Group 1 Vehicle  0 mg/kg Intravenous QD Days 1, 8, 15,22 1 21.4 4.4 22.4 9.3 Control Subcutaneous BID Days 2-7, 9-14, 2 21.37.6 21.9 10.6 16-20, 23-27 3 24.4 5.6 24.4 5.6 4 22.3 4.7 22.8 7.0 519.7 3.7 20.9 10.0 6 23.6 6.8 23.9 8.1 7 23.8 7.7 25.1 13.6 8 21.3 −1.422.1 2.3 9 21.8 −1.4 22.7 2.7 10 23.4 3.1 23.8 4.8 Group 2 PGX 48 mg/kgIntravenous QD Days 2, 8, 15, 22 1 23.1 4.5 24.3 10.0 ODSH 2 22.4 10.323.2 14.3 PGX 24 mg/kg Subcutaneous BID Days 2-7, 9-14, 3 22.0 4.8 22.88.6 ODSH 16-20 4 22.7 4.1 23.7 8.7 5 25.4 17.6 26.7 23.6 6 23.2 8.4 23.811.2 7 20.9 5.6 22.0 11.1 8 22.7 14.6 23.2 17.2 9 21.9 4.8 22.9 9.6 1020.9 6.1 21.9 11.2 Group 3 Carboplatin 80 mg/kg Intraperitoneal QD 1, 8,15 1 19.0 −6.4 19.7 −3.0 2 19.2 −11.1 19.3 −10.6 3 20.5 −7.7 20.1 −9.5 421.5 −7.3 22.7 −2.2 5 19.6 −4.9 20.8 1.0 6 22.8 0.9 23.2 2.7 7 20.5 −5.521.6 −0.5 8 21.2 2.4 22.7 9.7 9 21.6 −3.6 23.1 3.1 10 18.7 −5.1 19.4−1.5 Group 4 PGX 48 mg/kg Intravenous QD Days 1, 8, 15, 22 1 21.7 3.322.1 5.2 ODSH 2 21.9 −2.2 23.2 3.6 PGX 24 mg/kg Subcutaneous BID Days2-7, 9-14, 3 20.0 −7.4 20.5 −5.1 ODSH 16-20, 23-27 4 FD FD FD FDCarboplatin 80 mg/kg Intraperitoneal QD Days 1, 8, 15 5 22.9 4.6 23.99.1 6 20.7 1.0 21.4 4.4 7 18.3 −9.9 16.8 −17.2 8 22.1 −7.9 21.0 −12.5 921.7 2.8 23.3 10.4 10 24.2 −3.2 25.5 2.0 Individual weights (g) 7thWeight 8th Weight Day 18 Day 21 Weight Weight Weight change Weightchange Groups Compound Dosage Route Schedule Mouse (g) (%) (g) (%) Group1 Vehicle  0 mg/kg Intravenous QD Days 1, 8, 15, 22 1 22.6 10.2 23.012.2 Control Subcutaneous BID Days 2-7, 9-14, 2 22.6 14.1 23.3 17.716-20, 23-27 3 24.9 7.8 24.9 7.8 4 23.0 8.0 22.9 7.5 5 21.6 13.7 21.814.7 6 21.3 −3.6 23.1 4.5 7 24.7 11.8 24.2 9.5 8 22.2 2.8 22.6 4.6 922.3 0.9 23.1 4.5 10 24.3 7.0 23.8 4.8 Group 2 PGX 48 mg/kg IntravenousQD Days 2, 8, 15, 22 1 23.4 5.9 23.7 7.2 ODSH 2 23.2 14.3 24.0 18.2 PGX24 mg/kg Subcutaneous BID Days 2-7, 9-14, 3 23.3 11.0 23.2 10.5 ODSH16-20 4 23.2 6.4 24.0 10.1 5 26.4 22.2 26.4 22.2 6 23.8 11.2 24.1 12.6 720.0 1.0 20.4 3.0 8 21.8 10.1 21.9 10.6 9 23.0 10.0 22.9 9.6 10 21.2 7.621.9 11.2 Group 3 Carboplatin 80 mg/kg Intraperitoneal QD 1, 8, 15 118.7 −7.9 19.2 −5.4 2 17.9 −17.1 18.2 −15.7 3 20.2 −9.0 20.3 −8.6 4 22.2−4.3 23.1 −0.4 5 20.2 −1.9 21.4 3.9 6 21.3 −5.8 21.5 −4.9 7 19.4 −10.619.9 −8.3 8 22.0 6.3 23.5 13.5 9 23.3 4.0 24.0 7.1 10 18.0 −8.6 18.0−8.6 Group 4 PGX 48 mg/kg Intravenous QD Days 1, 8, 15, 22 1 21.2 1.020.7 −1.4 ODSH 2 22.5 0.4 22.5 0.4 PGX 24 mg/kg Subcutaneous BID Days2-7, 9-14, 3 19.4 −10.2 19.8 −8.3 ODSH 16-20, 23-27 4 FD FD FD FDCarboplatin 80 mg/kg Intraperitoneal QD Days 1, 8, 15 5 22.1 0.9 23.15.5 6 19.3 −9.9 19.1 −6.8 7 17.7 −12.8 18.1 −10.8 8 19.8 −17.5 20.7−13.8 9 21.9 3.8 22.6 7.1 10 23.3 −6.8 23.8 −4.8 FD = Found Dead TS =Terminal Sacrifice

Individual weights (g) 9th Weight 10th Weight Day 25 Day 28 WeightWeight Weight change Weight change Groups Compound Dosage Route ScheduleMouse (g) (%) (g) (%) Group 1 Vehicle  0 mg/kg Intravenous QD Days 1, 8,15, 22 1 22.5 9.8 24.0 17.1 Control Subcutaneous BID Days 2-7, 9-14, 2TS TS TS TS 16-20, 23-27 3 25.9 12.1 26.2 13.4 4 TS TS TS TS 5 TS TS TSTS 6 24.1 9.0 24.0 8.6 7 TS TS TS TS 8 23.9 10.6 20.7 −4.2 9 TS TS TS TS10 23.3 2.6 24.2 6.6 Group 2 PGX 48 mg/kg Intravenous QD Days 2, 8, 15,22 1 TS TS TS TS ODSH 2 24.3 19.7 29.0 23.2 PGX 24 mg/kg SubcutaneousBID Days 2-7, 9-14, 3 TS TS TS TS ODSH 16-20 4 TS TS TS TS 5 28.3 31.028.7 32.9 6 TS TS TS TS 7 21.7 9.6 22.9 15.7 8 23.4 18.2 23.8 20.2 924.0 14.8 24.2 15.8 10 TS TS TS TS Group 3 Carboplatin 80 mg/kgIntraperitoneal QD 1, 8, 15 1 19.9 −2.0 20.9 3.0 2 TS TS TS TS 3 20.4−8.1 20.3 −8.6 4 TS TS TS TS 5 22.3 8.3 23.0 11.7 6 21.8 −3.5 22.2 −1.87 20.2 −6.9 21.1 −2.8 8 TS TS TS TS 9 TS TS TS TS 10 TS TS TS TS Group 4PGX 48 mg/kg Intravenous QD Days 1, 8, 15, 22 1 22.9 9.0 22.9 9.0 ODSH 223.6 5.4 23.8 6.3 PGX 24 mg/kg Subcutaneous BID Days 2-7, 9-14, 3 TS TSTS TS ODSH 16-20, 23-27 4 FD FD FD FD Carboplatin 80 mg/kgIntraperitoneal QD Days 1, 8, 15 5 TS TS TS TS 6 TS TS TS TS 7 TS TS TSTS 8 TS TS TS TS 9 23.3 10.4 23.9 13.3 10 23.5 2.0 26.3 5.2 FD = FoundDead TS = Terminal Sacrifice

6.3. Example 3 ODSH, a PF4-Interacting Heparinoid, AttenuatesThrombocytopenia and Neutropenia, Induces Thrombopoiesis and NeutrophilProduction, and Reduces Constitutional Symptoms in Patients Receiving aChemotherapy Treatment Regimen Having Myelosuppressive Side Effects

Patients diagnosed with metastatic pancreatic cancer were treated withODSH as an adjunct to treatment with gemcitabine and nab-paclitaxel(Abraxane®, albumin-bound paclitaxel) in an unblinded clinical trial.

Inclusion criteria. Male and non-pregnant, non-lactating, femalepatients, aged 18 to 75, with histologically confirmed metastaticadenocarcinoma of the pancreas were enrolled in the trial. Furtherinclusion criteria were: presence of at least one metastatic tumors(measurable by conventional techniques or CT scan), serum CA 19-9measurement of greater than 2 times the upper limit of normal, noradiation therapy or chemotherapy for locally advanced disease withinsix months of enrollment into the trial, absolute neutrophil count of atleast 1.5×10⁹/L, platelet count of at least 100,000/mm³ (or 100×10⁹/L),hemoglobin level of at least 9 g/dL, prothrombin and partialthromboplastin times within normal limits (+/−15%) at time of screening,Eastern Cooperative Oncology Group performance status of 1 or more. Inaddition, patients were screened for blood chemistry levels, includingserum creatinine levels within normal limits, serum transaminase levelsof 2.5 or greater than the upper limit of normal, and bilirubin levelsof 1.5 times the upper limit of normal or more.

Dosing regimen. Treatment consisted of a series of 28-day cycles,wherein the last day of one cycle (e.g., day 28, cycle 1) wasimmediately followed by the first day of the next cycle (e.g., day 1,cycle 2). In each 28-day treatment cycle, patients were dosed at days 1,8, and 15 (three weeks of medication followed by one week of rest) asfollows. First, nab-paclitaxel at 125 mg/m² was administered as anintravenous infusion over 30 minutes. Next, gemcitabine was administeredat 1000 mg/m² as an intravenous infusion over 30 minutes. Nab-paclitaxeland gemcitabine therapy was given as described in the prescribinginformation for Abraxane® and Gemzar®. See also, Von Hoff et al., 2011,J. Clinical Oncology 29:1-7).

ODSH was administered immediately after gemcitabine administration, asfollows: an initial loading dose was administered as a bolus of 4 mg/kgover 5 minutes followed by a continuous intravenous infusion over 48hours, at a dose of 0.375 mg/kg/hr. These doses are, respectively,9-fold higher and 3.75-fold higher than the bolus and maintenance dosesof unfractionated heparin routinely used to confer full anticoagulationin a clinical setting (0.44 mg/kg for bolus injection, and 0.1 mg/kg/hrfor maintenance). As a substantially non-anticoagulant heparinoid, ODSHcan be administered at a dose that is greater than the fullyanticoagulant dose of unfractionated heparin, without concern foranticoagulant effect, as shown in the Results below.

An initial run-in period was conducted in which ten patients weretreated with the gemcitabine, nab-paclitaxel, and ODSH regimen describedabove. After all ten patients had completed at least one 28-day cycle,the data were reviewed and an open-label randomized study initiated,with two arms (ODSH arm and Control arm). In the ODSH arm, the dosingregimen was the same as in the run-in period (gemcitabine,nab-paclitaxel, and ODSH). In the Control arm, patients were given thesame gemcitabine and nab-paclitaxel regimen as in the run-in period, butwithout ODSH.

Testing. Blood was drawn before treatment administration on days 1, 8,and 15 of each 28 day cycle. Platelet counts, total white blood cellcounts, and absolute neutrophil counts were performed. Grading ofthrombocytopenia and/or neutropenia was performed according to thefollowing standards:

Platelet count Grade (×10³/μl blood) Thrombocytopenia  0* >150 1<150-75  2 <75-50 3 <50-25 4  <25 *(non-thrombocytopenic) Platelet countGrade (×10³/μl blood) Neutropenia  0* ≥2 1  <2-1.5 2 <1.5-1  3 <1.0-0.54   <0.5 *(non-neutropenic)Blood samples were also tested on day 1 of each treatment cycle for theserum level of CA19-9 (carbohydrate antigen 19-9) a marker used toassess the efficacy of the chemotherapy agents in treating pancreaticcancer. See, e.g., Maeda, 2011, Int. J. Clin. Oncol. 16(5):539-45.Reduction in the serum level of CA19-9 is indicative of chemotherapeuticefficacy against the pancreatic cancer. Tumor response and diseasestatus was measured according to Response Evaluation Criteria in SolidTumors (RECIST) Guidelines Version 1.1. Eur. J. Cancer, 2009,45:228-247. A reduction of at least 30% in the sum of the diameters oftarget lesions (measurable lesions present at screening, up to 2 lesionsper involved organ) was scored as a partial response. Patients werescored as having stable disease where the smallest sum of target lesiondiameters neither decreased nor increased sufficiently to qualify as apartial response or progressive disease.

Lack of anticoagulation by ODSH was confirmed by monitoring partialthromboplastin time (aPTT) during ODSH infusion, at days 1, 3, 10, and17 of treatment cycle 1. Normal range of aPTT is about 27 to 35 seconds+/−15%.

Patients were also assessed for other side effects of chemotherapy,including fatigue, sensory neuropathy, nausea, and vomiting, each ofwhich was graded according to severity. Grades were: O (normal), 1(mild), 2 (moderate), 3 (severe) and 4 (life-threatening).

Results. Platelet counts are shown in Table 16 below. Counts shown inthe “Screen” column are counts prior to entry into the clinical trialprotocol.

Ten out of 10 patients who completed the first cycle of chemotherapy andwho also had blood drawn at the beginning of the second cycle hadplatelet counts at the beginning of the second cycle that were greaterthan their platelet counts at the beginning of the first cycle, beforeany chemotherapy had been administered. Eight out of 9 patients whocompleted the second cycle of chemotherapy and who also had blood drawnat the beginning of the third cycle had platelet counts at the beginningof the third cycle that were greater than their platelet counts at thebeginning of the first cycle, before any chemotherapy had beenadministered. This trend was robust, continuing into subsequent cyclesof chemotherapy. For example, 9 out of 10 patients who completed thethird cycle of chemotherapy and who also had blood drawn at thebeginning of the fourth cycle had platelet counts at the beginning ofthe fourth cycle that were greater than their platelet counts at thebeginning of the first cycle, before any chemotherapy had beenadministered.

TABLE 16 Platelet Count (×10³/μl blood) Cycle 1 Cycle 2 Cycle 3 Cycle 4Cycle 5 Cycle 6 Patient Day Day Day Day Day Day Day Day Day Day Day DayDay Day Day Day Day Day ID Screen 1 8 15 1 8 15 1 8 15 1 8 15 1 8 15 1 815 2001 204 342 206 134 614 375 186 628 371 171 626 507 169 607 377 6002505 429 214 260 532 257 170 387 362 189 382 448 202 427 403 194 503 493164 6003 No 180 86 63 274 95 54 361 179 71 387 185 78 288 188 72 275 14872 data 6004 321 313 205 135 419 434 209 510 435 207 586 464 203 499 231132 288 384 169 6006 No 264 167 178 322 336 207 327 325 170 402 339 170351 288 169 370 321 162 data 6007 187 191 108 108 312 241 132 362 362163 292 355 140 266 241 153 294 207 128 7001 269 274 150 75 574 424 167655 388 188 478 468 163 389 325 109 489 336 102 7002 267 276 239 220 445357 186 418 223 171 667 193 149 275 192 8001 166 120 86 76 175 83 70 123119 63 184 129 106 94 58 114 85 54 9001 271 271 230 181 417 402 178 487355 172 406 188 339 512 378 193 371 313 338

FIG. 8 shows a plot of the platelet counts for individual patients asmeasured in samples taken before treatment on indicated days (D1=day 1,D8=day 8, and D15=day 15) of the indicated cycles (C1=cycle 1; C2=cycle2, etc.). There is a clear trend of increasing platelet count at thestart of each successive cycle of treatment, relative not only to theplatelet count after two weeks of treatment in a previous cycle, butalso relative to the platelet count at the start of the initial cycle oftreatment, i.e. before any treatment was administered. None of thepatients exhibited thrombocytopenia at the start of the second cycle andonly one of 8 patients exhibited thrombocytopenia at the start of thethird cycle.

Table 17 shows the percentage of patients with thrombocytopenia aftertwo doses (day 15, first cycle) or after five or more doses (day 15,second cycle or day 1, third cycle) of treatment with ODSH adjunctive togemcitabine and nab-paclitaxel. Also shown in Table 17, at row 3, arehistorical data, showing the percentages of patients with varying gradesof thrombocytopenia who had been treated with gemcitabine andnab-paclitaxel in the same amounts and on the same dosing schedule asdescribed herein, but without adjunctive administration of ODSH. Thedata presented in row 3 are reproduced from Table 3 of Von Hoff et al.,2011, J. Clinical Oncology 29:1-7, which provides the overall number andpercent of patients exhibiting selected adverse events throughout thetrial. Total number of patients in each category is shown inparentheses.

TABLE 17 Toxicity (% patients with thrombocytopenia of indicated grade)Treatment (no. Row of patients) Grade 0 Grade 1 Grade 2 Grade 3 Grade 41 Gemcitabine + 40 50 10 0 0 nab-paclitaxel + ODSH, first cycle, day 15(10) 2 Gemcitabine + 70 10 20 0 0 nab-paclitaxel + ODSH, second cycleday 15 (10) 3 Von Hoff et al. 9 42 21 19 9 Gemcitabine + nab-paclitaxel(44) (historical)

As shown above, after two doses of treatment, 40% of the patientstreated adjunctively with ODSH showed no thrombocytopenia, and 90% ofthe patients had no more than mild thrombocytopenia. After five doses oftreatment (day 15 of cycle 2), 7 out of 10 patients had nothrombocytopenia.

Table 18 shows the platelet counts for each patient at day 1 ofsuccessive treatment cycles (e.g., treatment cycle 1, 2, 3, etc.). Ascan be seen from the platelet counts at the beginning of cycles 2 and 3,even after receiving one or two full cycles (equal to three or sixdoses) of gemcitabine and nab-paclitaxel, only one of the patients wasthrombocytopenic, exhibiting mild (Grade 1) thrombocytopenia. All otherpatients showed platelet counts well above the lower limit of normal.This trend continued into Cycle 4 for patients 2001, 6002, 6003, 6004,6006, 6007, 7001, 8001, and 9001. Overall, platelets counts in 10/10patients were higher at the beginning of cycle 2 than at the beginningof cycle 1, and platelets counts in 6/9 patients were higher at thebeginning of cycle 3 than at the beginning of cycle 2.

TABLE 18 Platelet count at start of cycle Patient Cycle number 1 Cycle 2Cycle 3 Cycle 4 Cycle 5 Cycle 6 Cycle 7 2001 342 614 628 626 6002 429532 387 382 427 503 6003 180 274 361 387 288 275 6004 313 419 510 586499 288 6006 264 322 327 402 351 6007 191 312 362 292 7001 274 574 655478 389 489 454 7002 276 445 418 667 275 8001 120 175 123 184 9001 271417 406 512 371

As shown further in FIG. 10A, mean and median platelet counts across allsamples were consistently above 150,000 on the first day of treatmentcycles 2 and 3, even when numbers were below 150,000 on day 15 of thefirst treatment cycle. In fact, in treatment cycles 2 and 3, mean andmedian platelet counts at day 15—when platelet counts are expected to beat their lowest—remained in the normal range. Furthermore, mean andmedian platelet counts at the start of the third cycle were greater thanat the start of the second cycle and were greater at the start of thesecond cycle than at the screening measurement and the start of thefirst cycle (before any chemotherapy was administered).

In the randomized stage of the trial, a statistically significantdifference in platelet counts was detected between the ODSH arm and theControl arm of the study, as shown in Table 19 below, demonstrating thatODSH attenuates thrombocytopenia associated with myelosuppressivetreatment regimens.

TABLE 19 Patient Cycle 1 Cycle 2 Cycle 3 Cycle 4 number Day 15 Day 15Day 15 Day 15 ODSH ARM 6009 199 92 96 138 8002 172 165 8005 162 9002 2329004 231 110 124 11001 216 156 14001 121 95 74 31 14002 103 74 76CONTROL ARM 4002 64 6008 92 81 38 8003 186 200 9003 71 119 53 78 1100277 14003 95 80 94 14005 104 118 117 One tail t-test Two tail t-testEffect on platelets .021 .042

Table 20 below provides absolute neutrophil counts for the patientstreated adjunctively with ODSH. Eight out of 10 patients who completedthe first cycle of treatment showed normal neutrophil counts on thefirst day of the second treatment cycle. Five out of 10 patients whocompleted the first cycle also had increased or unchanged neutrophilcounts on the first day of the second treatment cycle as compared to thefirst day of the first cycle (prior to chemotherapy). Furthermore, 8 outof 9 patients showed normal neutrophil counts at the start of the thirdcycle of treatment and 10 out of 10 patients showed normal neutrophilcounts at the start of the fourth cycle of treatment.

TABLE 20 Neutrophil Values (×10³/μl blood) Cycle 1 Cycle 2 Cycle 3 Cycle4 Cycle 5 Cycle 6 Patient Day Day Day Day Day Day Day Day Day Day DayDay Day Day Day Day ID Screen Day 1 8 15 Day 1 8 15 1 8 15 1 8 15 1 8 151 8 15 2001 9.3 4.0 1.0 1.3 3.4 2.3 2.7 3.1 1.8 2.6 3.8 6002 7.6 6.2 4.03.9 8.3 1.7 2.4 3.4 1.5 1.5 4.4 2.6 2.9 5.0 2.1 2.3 5.2 3.7 1.2 6003 No4.5 2.2 0.9 1.9 1.8 1.5 3.2 1.5 2.7 1.5 1.1 2.0 0.9 1.0 2.2 0.9 1.1 data6004 5.6 4.3 1.3 0.7 7.3 3.8 1.8 2.7 1.6 1.5 2.2 2.5 4.6 1.6 2.1 0.813.5 1.3 1.7 6006 No 3.6 2.1 1.6 1.6 2.6 1.7 2.8 4.2 1.8 5.2 2.7 2.4 3.23.4 3.9 data 6007 5.9 5.5 4.3 3.6 4.6 3.7 3.8 2.2 6.8 4.5 5.7 7001 4.03.6 0.5 0.7 13.0 6.3 0.7 3.9 3.9 1.4 5.9 2.2 3.4 3.3 1.6 1.1 2.3 1.210.7 7002 4.0 2.1 3.0 3.5 2.1 3.7 1.5 1.8 1.4 1.5 3.5 2.0 0.7 23.2 4.18001 5.3 3.3 1.5 1.0 6.0 1.3 1.1 3.2 1.4 0.8 3.2 9001 4.3 4.3 3.6 1.91.0 2.7 1.4 1.7

FIG. 9 shows a plot of the neutrophil count for individual patients asmeasured in samples taken before treatment on indicated days (D1=day 1,D8=day 8, and D15=day 15) of the indicated cycles (C1=cycle 1; C2=cycle2, etc.). There is a clear trend of increasing neutrophil count at thestart of the second cycle of treatment, relative to the neutrophil countat the start of the initial cycle of treatment, i.e. before anytreatment was administered.

Table 21 shows the percentage of patients with neutropenia after twodoses (day 15, first cycle) or after five or more doses (day 15, secondcycle or day 15, third cycle) of treatment with ODSH adjunctive togemcitabine and nab-paclitaxel. Also shown in Table 21, at row 3, arehistorical data showing the percentages of patients with varying gradesof neutropenia who had been treated with gemcitabine and nab-paclitaxelin the same amounts and on the same dosing schedule as described herein,without ODSH. The data presented in row 3 are reproduced from Table 3 ofVon Hoff et al., 2011, J. Clinical Oncology 29:1-7, which provides theoverall number and percent of patients exhibiting selected adverseevents throughout the trial. The total number of patients in eachcategory is shown in parentheses.

TABLE 21 Toxicity (% patients with neutropenia of indicated grade)Treatment (no. of Row patients) Grade 0 Grade 1 Grade 2 Grade 3 Grade 41 Gemcitabine + 33 11 22 33 0 nab-paclitaxel + ODSH, first cycle day 15(9) 2 Gemcitabine + 20 50 20 10 0 nab-paclitaxel + ODSH, second cycleday 15 or third cycle day 15 (10) 3 Von Hoff et al. 9 14 2 26 49Gemcitabine + nab-paclitaxel (44)

As shown above, after two doses of treatment, 44% of the patientstreated adjunctively with ODSH showed no more than mild neutropenia.After five doses of treatment (day 15 of cycle 2) or eight doses oftreatment (day 15 of cycle 3) 7 out of 10 patients had no more than mildneutropenia. As shown further in FIG. 10B, mean and median absoluteneutrophil counts across all samples, were consistently above 2,000 onthe first day of treatment cycles 2 and 3, even when numbers were below2,000 on day 15 of the previous cycle. With successive cycles andwithout dose reduction, patients with grade 3 neutropenia at the nadirof the first cycle, had no more than grade 1 or 2 neutropenia in cycles2 or 3 (see patients 6004, 6004, and 7001). Only one patient (7001) outof 10 was treated with G-CSF, receiving a single dose in cycle 1.

In the randomized stage of the trial, a statistically significantdifference in neutrophil counts was detected between the ODSH arm andthe Control arm of the study, as shown in Table 22 below, demonstratingthat ODSH attenuates neutropenia associated with myelosuppressivetreatment regimens.

TABLE 22 Patient Cycle 1 Cycle 2 Cycle 3 Cycle 4 number Day 15 Day 15Day 15 Day 15 ODSH Arm 6009 4.7 3.7 5.4 4.1 8002 3.0 3.7 8005 0.3 90024.0 9004 2.4 1.8 1.0 11001 1.4 1.8 14001 4.0 1.6 1.6 4.8 14002 0.8 0.80.8 Control Arm 4002 0.6 6008 1.1 1.1 0.3 8003 1.7 1.7 9003 0.8 0.8 1.00.9 11002 1.4 14003 3.3 2.2 3.1 14005 2.3 1.8 1.3 One tail t-test Twotail t-test Effect on neutrophils .009 .017

Total white blood cell counts were consistent with neutrophil counts, asshown below in Table 23.

TABLE 23 White Blood Cell Counts (×10³/μl blood) Cycle 1 Cycle 2 Cycle 3Cycle 4 Patient ID Screen Day 1 Day 8 Day 15 Day 1 Day 8 Day 15 Day 1Day 8 Day 15 Day 1 Day 8 Day 15 2001 10.8 5.7 2.3 2.7 5.7 4.5 4.3 6.06002 10.6 8.7 5.7 5.4 11.0 4.6 5.4 7.6 4.8 4.3 8.5 6.0 6003 6.0 6.0 3.41.5 3.4 2.9 2.4 4.8 2.7 6004 7.3 7.2 4.1 2.7 10.8 6.4 4.4 5.8 6006 Nodata 6.8 4.4 4.2 4.9 6007 8.5 7.7 6.2 5.1 7.8 7001 6.7 7.3 2.7 2.1 17.810.2 3.4 7002 6.3 5.0 5.3 6.0 5.9 8001 6.7 4.6 2.8 2.0 9001 6.5 6.5 5.13.2 6.5

Assessment of fatigue, sensory neuropathy, nausea, and vomiting, all ofwhich are common side effects of chemotherapy, and notably in thegemcitabine-abraxane regimen described in Von Hoff et al., revealed thatpatients treated adjunctively with ODSH experienced mild to moderatesymptoms, with more than 50% of the patients experiencing no more thanGrade 1 side effects. Table 24 below provides the percentage of patientsexperiencing different grades of each side effect, as compared to thepercentages reported in Von Hoff et al., 2011, J. Clinical Oncology29:1-7.

TABLE 24 Toxicity (% patients Side Treatment (no. per indicated grade)Effect of patients) Grade 0 Grade 1 Grade 2 Grade 3 Fatigue Von Hoff etal. 20 23 30 27 (44) Gemcitabine + 45 55 0 0 nab-paclitaxel + ODSH (9)Nausea Von Hoff et al. 53 25 20 2 (44) Gemcitabine + 45 55 0 0nab-paclitaxel + ODSH (9) Vomiting Von Hoff et al. 63 23 7 7 (44)Gemcitabine + 91 9 0 0 nab-paclitaxel + ODSH (9) Sensory Von Hoff et al.26 34 20 20 Neuropathy (44) Gemcitabine + 90 0 0 10 nab-paclitaxel +ODSH (9)

Serum CA19-9 level, a marker correlated with extent of tumor, andtherefore correlated with efficacy of the chemotherapeutic treatment,decreased in 8 of 10 patients, showing that the ability of ODSH toattenuate thrombocytopenia and neutropenia and induce thrombopoiesis andneutrophil production did not interfere with the efficacy of thechemotherapy (indicated by the decrease in CA19-9 serum levels),consistent with the results obtained in the pancreatic xenograft animalmodel described in Example 1. See Table 25 below.

TABLE 25 CA19-9 Serum levels (U/mL) Cycle 1 Cycle 2 Cycle 3 Cycle 4Patient ID Day 1 Day 1 Day 1 Day 1 2001 4,775 5,934 8,529 9,926 6002 12571 27 15 6003 6,186 4,392 2,669 2405 6004 6,275 4,018 1,433 348 6006 395490 251 108 6007 70,086 22,958 10,286 7001 2,138 502 328 288 7002 483453 205 110 8001 11 17 15 20 9001 326 710 164 69

All ten patients showed a response to treatment despite extensivemetastatic disease at the start of the clinical trial: 5 patients showeda partial response and 5 patients showed stable disease, as measuredusing RECIST Criteria. See FIG. 12A. After three or four full cycles oftreatment, no patient showed any clinical or radiographic evidence ofprogressive disease. Patients 2001 and 6002 also exhibited reduction inthe size of liver and lung or nodal metastases by the fourth cycle oftreatment. As shown in FIG. 11A, Liver metastases in patient 2001 havedisappeared and pulmonary lesions have decreased in size, with minimalclinical symptoms of metastatic disease, despite rising CA19-9 levels.Patient 6002 showed stable disease and a reduction in the size ofmetastatic lesions in the liver and lymph nodes. See FIGS. 11B-C.Patient 6003, who presented with lung metastases, also showed stabledisease, as shown in FIG. 11D. Patient 6006 showed stable disease andsome reduction in the size pancreatic and metastatic lesions in theliver. See FIG. 11E. Patient 8001 showed stable disease with a reductionin the size of pancreatic tumor at the end of cycle 2. See FIG. 11F.FIGS. 12B-F show that patients 6004, 6007, 7001, 7002, and 9001 had apartial response. Eastern Cooperative Oncology Group (ECOG) Performancestatus in 7 evaluable patients was stable or improved after 8 weeks inthe trial (at day 1 of treatment cycle 3).

The treatment regimen also appears to have minimal adverse effects onweight, as most patients have experienced minimum weight loss or evensome weight gain. See Table 26 below.

TABLE 26 Weight (in pounds) Cycle 1 Cycle 2 Cycle 3 Cycle 4 Cycle 5Cycle 6 Patient ID Day 1 Day 1 Day 1 Day 1 Day 1 Day 1 2001 125 124 119120 6002 163 164 169 173 177 176 6003 147 140 144 139 137 133 6004 158155 157 153 156 157 6006 189 187 188 185 188 6007 110 115 112 115 7001163 164 162 164 163 7002 197 191 186 170 167 8001 160 164 166 166 9001137 135 134 129 127

As shown in Table 27 below, ODSH infusion did not result inanticoagulation, as determined by partial thromboplastin time (aPTT) inthe patients.

TABLE 27 aPTT (in seconds) Cycle 1 Cycle 1 Cycle 1 Cycle 1 Patient IDDay 1 Day 3 Day 10 Day 17 2001 26 34 33 33 6002 25 31 36 6003 29 33 346004 27 31 31 29 6006 29 34 34 6007 24 28 28 7001 33 32 30 7002 36 43 3842 8001 33 38 35 36 9001 28 31 31 30

In the open-label randomized trial initiated after the run-in period, asignificant difference in platelet count was observed between the ODSHarm and the Control arm (no ODSH), as shown in FIGS. 13A and 13B. Themedian and mean platelet counts at day 15 of the first cycle ofchemotherapy (after three doses of chemotherapy) were significantlyhigher in the patients receiving ODSH in addition to gemcitabine andnab-paclitaxel than in the patients not receiving ODSH (p =0.013 usingunpaired t-test, 5 patients in each treatment arm). See FIG. 13A. Thiseffect held true in subsequent cycles 2 through 6 (p =0.0003 after 6cycles in unpaired t-test). See FIG. 13B and Table 28 below.Furthermore, adjunctive administration of ODSH enhanced plateletrecovery by day 1 of a subsequent cycle. FIG. 13B and Table 28 below(compare day 1 of cycles in control arm patients versus ODSH armpatients, p =0.0004 after 6 cycles, unpaired t-test).

TABLE 28 Day 1 Day 15 Day 1 Day 15 (n) (n) CONTROL Cycle 1 223 95 16 13ARM Cycle 2 331 107 11 9 Cycle 3 286 95 9 8 Cycle 4 242 101 7 7 Cycle 5255 85 6 6 Cycle 6 257 50 6 5 ODSH ARM Cycle 1 248 143 26 26 Cycle 2 336135 24 21 Cycle 3 393 171 14 13 Cycle 4 404 156 12 12 Cycle 5 389 121 118 Cycle 6 371 132 8 8

In conclusion, ODSH attenuated the myelosuppressive side effects of thegemcitabine/Abraxane regimen (as compared to patients reported in VonHoff et al., 2011, J. Clinical Oncology 29:1-7 and as compared topatients randomized to receive gemcitabine/Abraxane alone, in therandomized portion of this trial), and increased platelet and neutrophilcounts in the patients above levels seen prior to treatment, whilepreserving the efficacy of the chemotherapy regimen. Furthermore, areduction in side effects which manifest as constitutional symptoms,such as fatigue, nausea, and vomiting, was also observed. ODSH and otherPF4-interacting heparinoids attenuate constitutional symptoms associatedwith chemotherapy.

Overall, the observed effects may permit intensification of thechemotherapeutic regimen with improved antineoplastic efficacy.

6.4. Example 4 ODSH Attenuates Thrombocytopenia Associated with theTreatment of Acute Myelogenous Leukemia (AML)

A clinical trial is conducted to confirm the therapeutic advantage ofODSH administered adjunctively to induction and consolidation therapyand subsequent allogeneic or autologous bone marrow transplantation inthe treatment of acute myelogenous leukemia (AML). Subjects included inthe trial are subjects diagnosed with AML who are undergoing inductionand consolidation therapy. Subjects are randomly assigned to either acontrol group (receiving only induction and consolidation therapy) or atreatment group (receiving adjunctive administration of ODSH). ODSH isadministered as a continuous infusion (0.375 mg/kg/hr). Subjects in eacharm of the trial are evaluated for platelet counts and the need forplatelet transfusion. Further metrics evaluated include measurement ofcirculating levels of PF4 and rate of granulocyte recovery.

Results are obtained which demonstrate that addition of ODSH to standardinduction and consolidation therapy attenuates thrombocytopenia.

A second clinical trial is conducted to confirm the advantage of ODSHadministered adjunctively to induction and consolidation chemotherapy inthe treatment of AML. The trial is an open-label pilot study of tenpatients newly diagnosed with AML and not previously treated for AML.Patient treatment regimens are as follows. During an induction phase,100 mg/m²/day of cytarabine is administered continuously by intravenousinfusion for 7 days (Day 1-Day 7) and 12 mg/m²/day of idarubicin isadministered by intravenous injection on each of Day 1, Day 2, and Day3.Four mg/kg ODSH is administered intravenously as a bolus on Day 1immediately after idarubicin, and is then administered at a dose of 0.25mg/kg/hr continuously by intravenous infusion on Day 1 to Day7. Forpatients under 60 years of age, the induction phase is followed by up tofour cycles of consolidation chemotherapy, each cycle consisting of 3g/m² cytarabine administered over a period of 3 hours, every 12 hours onDays 1, 3, and 5 or a 5-day cycle. During the consolidation phase, 4mg/kg ODSH is administered intravenously as a bolus on Day 1 immediatelyafter cytarabine, and is then administered at a dose of 0.25 mg/kg/hrcontinuously by intravenous infusion on Day 1 to Day 5. Consolidationchemotherapy is initiated no sooner than 28 days from the start ofinduction chemotherapy. Subjects are evaluated for the degree andduration of thrombocytopenia, based on platelet counts and need fortransfusion.

Results are obtained which demonstrate that addition of ODSH to standardinduction and consolidation therapy attenuates thrombocytopenia.

All publications, patents, patent applications and other documents citedin this application are hereby incorporated by reference in theirentireties for all purposes to the same extent as if each individualpublication, patent, patent application or other document wereindividually indicated to be incorporated by reference for all purposes.

While various specific embodiments have been illustrated and described,it will be appreciated that various changes can be made withoutdeparting from the spirit and scope of the invention(s).

What is claimed is:
 1. A method of attenuating a myelosuppressive sideeffect of a patient treatment regimen comprising: adjunctivelyadministering a therapeutically effective amount of a PF4-interactingheparinoid to the patient.